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Background: Recent anatomical studies have reported the feasibility of the endoscopic endonasal approach to the middle fossa. However, its clinical applicability has been discussed in only a few cases. This article describes the case of a middle fossa epidural abscess successfully drained through a fully endoscopic endonasal corridor and discusses the key technical points., Observations: The authors describe an 8-year-old boy who presented with worsening headache, fatigue, emesis, and fever and was diagnosed with a left middle fossa epidural abscess associated with sphenoid sinusitis. Following endoscopic sinus surgery performed by a rhinologist, the middle fossa was accessed by removing the maxillary strut through the transmaxillary transpterygoid corridor. Complete drainage of the abscess was confirmed on postoperative magnetic resonance imaging. The patient tolerated the surgery without neurological deficit and demonstrated prompt symptom improvement. He was discharged home after completing a 6-week course of antibiotic therapy and remained free from recurrence at 1 year following surgery., Lessons: The endoscopic endonasal approach may be applicable to a middle fossa epidural abscess, resulting in prompt clinical improvement. The maxillary strut is a key structure for entering the middle fossa. https://thejns.org/doi/10.3171/CASE24288.

Background: There is no clear objective indicator for selecting soft foods that are required for food bolus formation in older people with impaired oral function., Objective: This study aimed to investigate the relationship between maximal isometric tongue pressure (MITP) and the mechanical properties of gels that can be crushed by the tongue., Methods: This study included 65 healthy participants aged 22-96 (young group; 15 males, 15 females; older dentate group; 7 males, 8 females; older edentulous group; 10 males, 10 females). MITP was measured by the balloon-probe device. Agar gel with 10 different kinds of fracture force from 10N to 100N was used. The limit of fracture force of gels (LFFG) that were crushed by the tongue was measured by the up-and-down method. In the older edentulous group, two items were measured with and without dentures. Spearman's rank correlation coefficient was used to evaluate the relationship between MITP and LFFG in each group (p < .05)., Results: There were positive correlations between MITP and LFFG in all groups (overall groups: r s  = .66, young group: r s  = .46, older dentate group: r s  = .61, older edentulous group with dentures: r s  = .60, older edentulous group without dentures: r s  = .47)., Conclusion: MITP and LFFG were positively correlated in young, older dentate and older edentulous groups, suggesting that MITP has the potential to be an objective indicator of the range of mechanical properties of soft food that can be crushed by the tongue., (© 2023 John Wiley & Sons Ltd.)

Background and Aims: To examine the roles of microRNAs in the development of colitis, we conducted the RNA-sequencing studies using RNA derived from normal and colitogenic CD4+ T cells. Colitogenic CD4 + T cells demonstrated the increased expression of miR-150. We focused on the involvement of miR-150 in the colitis., Methods: We crossed miR-150 knockout mice and T-cell-specific Rap1KO mice, which is colitis model mice and spontaneously develop the colitis with tubular adenomas in microbiota-dependent manner., Results: MiR-150 silencing completely inhibited the expansion of pathogenic Th17 cells and the development of colitis., Conclusion: MiR-150 is a potential therapeutic target of inflammatory bowel diseases., (© 2023 The Authors.)

Background: Subjective-physiological emotional coherence is thought to be associated with enhanced well-being, and a relationship between subjective-physiological emotional coherence and superior nutritional status has been suggested in older populations. However, no study has examined subjective-physiological emotional coherence among older adults while tasting food. Accordingly, the present study compared subjective-physiological emotional coherence during food consumption among older and younger adults., Methods: Participants consumed bite-sized gel-type foods with different flavors and provided their subjective ratings of the foods while their physiological responses (facial electromyography (EMG) of the corrugator supercilia, masseter, and suprahyoid, and other autonomic nervous system signals) were simultaneously measured., Results: Our primary findings were that (1) the ratings of liking, wanting, and valence were negatively correlated with corrugator EMG activity in older and young adult participants; (2) the positive association between masseter EMG activity and ratings of wanting/valence was weaker in the older than in the young adult group; and (3) arousal ratings were negatively correlated with corrugator EMG activity in the older group only., Conclusions: These results demonstrate commonalities and differences in subjective-physiological emotional coherence during food intake between older and young adults.

Background: Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) and mild encephalopathy associated with excitotoxicity (MEEX) are the most frequent acute encephalopathies in pediatric patients in Japan. AESD typically presents with biphasic seizures and delayed reduced diffusion in the subcortical area, called bright tree appearance (BTA), on radiological examination. In patients with AESD, arterial spin labeling (ASL) shows decreased cerebral blood flow (CBF) in the hyperacute stage and increased CBF in the acute stage, suggesting the usefulness of ASL for the early diagnosis of AESD. Additionally, proton magnetic resonance spectroscopy (MRS) shows elevated glutamate (Glu) and glutamine (Gln) in AESD. MEEX is a group of mild encephalopathies with transient elevation of Gln on MRS similar to that in AESD; however, MEEX does not include any clinical biphasic course or abnormalities, including BTA on diffusion-weighted imaging. Although the usefulness of ASL for AESD has been reported, there are no reports for patients with MEEX. In this study, we report our experience with a 4-year-old girl diagnosed with MEEX who showed unique findings on ASL., Case Presentation: The patient was a 4-year-old girl admitted to the emergency room with febrile status epilepticus. Considering the possibility of AESD, vitamin therapy was initiated. ASL-MR imaging (MRI) of the brain performed on the second day showed increased blood flow in the frontal, temporal, and occipital regions with spared central sulcus, which indicated AESD with central sparing. The patient was diagnosed with AESD, and the treatment included pulse steroid therapy and immunoglobulin therapy from day 3. The patient remained mildly unconscious but gradually became conscious by day 7 with no seizures. Brain MRI performed on day 8 did not show any characteristic AESD findings, such as BTA. Furthermore, MRS showed elevated Gln, which, along with the clinical course, led to the diagnosis of MEEX. The patient was discharged on day 16 without obvious sequelae., Conclusions: ASL may be useful in the early diagnosis of MEEX as well as AESD, facilitating early intervention., (© 2022. The Author(s).)

We present a pattern-recognition-based sensor that targets cell-derived components in culture media and evaluates cultured cells without damaging them. An array sensor made of a single-polymer probe was employed to obtain fluorescence response patterns of the analyte media, allowing successful identification of the type and state of the cells via multivariate analysis.

The development of non-invasive techniques to characterize cultured cells is invaluable not only to ensure the reproducibility of cell research, but also for quality assurance of industrial cell products for purposes such as regenerative medicine. Here, we present a polymer-based 'chemical tongue', i.e. , a biosensing technique that mimics the human taste system, that is capable of non-invasively generating fluorescence response patterns that reflect the proteins secreted, and also partially consumed, by cultured cells, even from serum-supplemented media containing abundant interferants. Analysis of the spent media collected during cell culture using our chemical tongue, which consists of cationic polymers of different scaffolds appended with environmentally responsive dansyl fluorophores, led to the successful (i) identification of human-derived cell lines, (ii) monitoring of the differentiation process of stem cells, even at the stage when conventional staining was negative, and (iii) detection of cancer-cell contamination in stem cells. Since the characterization of cultured cells is usually performed via invasive methods that result in cell death, our chemical-tongue approach, which is of high practical utility, will offer a new means of addressing the growing demand for highly controlled cell production in the medical and environmental fields.

Gut-microbiota analysis has been recognized as crucial in health management and disease treatment. Metagenomics, a current standard examination method for the gut microbiome, is effective but requires both expertise and significant amounts of general resources. Here, we show highly accessible sensing systems based on the so-called chemical-nose strategy to transduce the characteristics of microbiota into fluorescence patterns. The fluorescence patterns, generated by twelve block copolymers with aggregation-induced emission (AIE) units, were analyzed using pattern-recognition algorithms, which identified 16 intestinal bacterial strains in a way that correlates with their genome-based taxonomic classification. Importantly, the chemical noses classified artificial models of obesity-associated gut microbiota, and further succeeded in detecting sleep disorder in mice through comparative analysis of normal and abnormal mouse gut microbiota. Our techniques thus allow analyzing complex bacterial samples far more quickly, simply, and inexpensively than common metagenome-based methods, which offers a powerful and complementary tool for the practical analysis of the gut microbiome., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

T-cell-specific Rap1 deletion causes spontaneous colitis in mice. In the present study, we revealed that Rap1 deficiency in T cells impaired the preceding induction of intestinal RORγt + Treg cells. In the large intestinal lamina propria (LILP) of T-cell-specific Rap1-knockout mice (Rap1KO mice), Th17 cells were found to increase in a microbiota-dependent manner, and the inhibition of IL-17A production prevented the development of colitis. In the LILP of Rap1KO mice, RORγt + Treg cells were scarcely induced by 4 weeks of age. The expression of CTLA-4 on Rap1-deficient Treg cells was reduced and the expression of CD80 and CD86 on dendritic cells was consequently elevated in Rap1KO mice. When cultured under each polarizing condition, Rap1-deficient naïve CD4 + T cells did not show biased differentiation into Th17 cells; their differentiation into Treg cells as well as Th1 and Th2 cells was lesser than that of wild-type cells. Rap1-deficient naïve CD4 + T cells were found to exhibit the defective nuclear translocation of NFAT and formation of actin foci in response to TCR engagement. These data suggest that Rap1 amplifies the TCR signaling required for Treg-mediated control of intestinal colitogenic Th17 responses., (© 2022. The Author(s).)

Sensing subjective hedonic or emotional experiences during eating using physiological activity is practically and theoretically important. A recent psychophysiological study has reported that facial electromyography (EMG) measured from the corrugator supercilii muscles was negatively associated with hedonic ratings, including liking, wanting, and valence, during the consumption of solid foods. However, the study protocol prevented participants from natural mastication (crushing of food between the teeth) during physiological data acquisition, which could hide associations between hedonic experiences and masticatory muscle activity during natural eating. We investigated this issue by assessing participants' subjective ratings (liking, wanting, valence, and arousal) and recording physiological measures, including EMG of the corrugator supercilii, zygomatic major, masseter, and suprahyoid muscles while they consumed gel-type solid foods (water-based gellan gum jellies) of diverse flavors. Ratings of liking, wanting, and valence were negatively correlated with corrugator supercilii EMG and positively correlated with masseter and suprahyoid EMG. These findings imply that subjective hedonic experiences during food consumption can be sensed using EMG signals from the brow and masticatory muscles.

Navigation of fast migrating cells such as amoeba Dictyostelium and immune cells are tightly associated with their morphologies that range from steady polarized forms that support high directionality to those more complex and variable when making frequent turns. Model simulations are essential for quantitative understanding of these features and their origins, however systematic comparisons with real data are underdeveloped. Here, by employing deep-learning-based feature extraction combined with phase-field modeling framework, we show that a low dimensional feature space for 2D migrating cell morphologies obtained from the shape stereotype of keratocytes, Dictyostelium and neutrophils can be fully mapped by an interlinked signaling network of cell-polarization and protrusion dynamics. Our analysis links the data-driven shape analysis to the underlying causalities by identifying key parameters critical for migratory morphologies both normal and aberrant under genetic and pharmacological perturbations. The results underscore the importance of deciphering self-organizing states and their interplay when characterizing morphological phenotypes., Competing Interests: The authors have declared that no competing interests exist.

Liquid-liquid phase separation (LLPS) of proteins and DNAs has been recognized as a fundamental mechanism for the formation of intracellular biomolecular condensates. Here, we show the role of the constituent DNA components, i.e., the phosphate groups, deoxyribose sugars, and nucleobases, in LLPS with a polycationic peptide, linker histone H1, a known key regulator of chromatin condensation. A comparison of the phase behavior of mixtures of H1 and single-stranded DNA-based oligomers in which one or more of the constituent moieties of DNA were removed demonstrated that not only the electrostatic interactions between the anionic phosphate groups of the oligomers and the cationic residues of H1, but also the interactions involving nucleobases and deoxyriboses (i) promoted the generation of spherical liquid droplets via LLPS as well as (ii) increased the density of DNA and decreased its fluidity within the droplets under low-salt conditions. Furthermore, we found the formation of non-spherical assemblies with both mobile and immobile fractions at relatively higher concentrations of H1 for all the oligomers. The roles of the DNA components that promote phase separation and modulate droplet characteristics revealed in this study will facilitate our understanding of the formation processes of the various biomolecular condensates containing nucleic acids, such as chromatin organization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mimura, Tomita, Sugai, Shinkai, Ishihara and Kurita.)

Integrin regulation by Rap1 is indispensable for lymphocyte recirculation. In mice having B-cell-specific Rap1a/b double knockouts (DKO), the number of B cells in lymph nodes decreased to approximately 4% of that of control mice, and B cells were present in the spleen and blood. Upon the immunization with NP-CGG, DKO mice demonstrated the defective GC formation in the spleen, and the reduced NP-specific antibody production. In vitro , Rap1 deficiency impaired the movement of activated B cells along the gradients of chemoattractants known to be critical for their localization in the follicles. Furthermore, B-1a cells were almost completely absent in the peritoneal cavity, spleen and blood of adult DKO mice, and the number of B-cell progenitor/precursor (B-p) were reduced in neonatal and fetal livers. However, DKO B-ps normally proliferated, and differentiated into IgM + cells in the presence of IL-7. CXCL12-dependent migration of B-ps on the VCAM-1 was severely impaired by Rap1 deficiency. Immunostaining study of fetal livers revealed defects in the co-localization of DKO B-ps and IL-7-producing stromal cells. This study proposes that the profound effects of Rap1-deficiency on humoral responses and B-1a cell generation may be due to or in part caused by impairments of the chemoattractant-dependent positioning and the contact with stromal cells., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ishihara, Sato, Sugioka, Miwa, Saito, Sato, Fukuyama, Nakajima, Sawai, Kotani and Katagiri.)

To provide appropriate foods for elderly people with eating difficulties, it is necessary to take account of the ability of compensatory mastication such as tongue squeezing. However, the biomechanics of tongue squeezing is still unclear. The aim of present study is to investigate the effect of the initial mechanical properties of gels on the change in tongue pressure production during squeezing and swallowing. As test sample, nine kinds of gels with three fracture force and three fracture strain were prepared. Tongue pressure during squeezing and swallowing gels was measured by using an ultra-thin tongue pressure sensor with five measuring points attached on the hard palate in seven healthy participants, and analyzed at four phases; Initial squeeze, Middle squeeze, Last squeeze, and Swallowing. The maximal magnitude of tongue pressure was increased for gels with higher fracture force at most measuring points and was decreased for gels with higher fracture strain at some measuring points on the median line during Initial and Middle squeezing. However, no influence by fracture force and strain was found in magnitude during Last squeezing and Swallowing. The duration of tongue pressure increased for gels with higher force at most measuring points during Middle squeezing, although no influence by strain was found during each phase. The results clearly show how the initial fracture properties of gels influence on tongue pressure production during each phase of food oral processing, which clarified one aspect of squeezing with tongue, as the compensatory mastication., (© 2021 Wiley Periodicals LLC.)

The development of a versatile sensing strategy for the damage-free characterization of cultured cells is of great importance for both fundamental biological research and industrial applications. Here, we present a pattern-recognition-based cell-sensing approach using a multichannel surface plasmon resonance (SPR) chip. The chip, in which five cysteine derivatives with different structures are immobilized on Au films, is capable of generating five unique SPR sensorgrams for the cell-secreted molecules that are contained in cell culture media. An automatic statistical program was built to acquire kinetic parameters from the SPR sensorgrams and to select optimal parameters as "pattern information" for subsequent multivariate analysis. Our system rapidly (∼10 min) provides the complex information by merely depositing a small amount of cell culture media (∼25 μL) onto the chip, and the amount of information obtained is comparable to that furnished by a combination of conventional laborious biochemical assays. This noninvasive pattern-recognition-based cell-sensing approach could potentially be employed as a versatile tool for characterizing cells.

Integrin activation is associated with conformational regulation. In this study, we developed a system to evaluate conformational changes in α 4 β 7 integrin. We first inserted the PA tag into the plexin-semaphorin-integrin (PSI) domain of β 7 chain, which reacted with an anti-PA tag antibody (NZ-1) in an Mn 2+ -dependent manner. The small GTPase Rap1 deficiency, as well as chemokine stimulation and the introduction of the active form of Rap1, Rap1V12, enhanced the binding of NZ-1 to the PA-tagged mutant integrin, and increased the binding affinity to mucosal addressing cell adhesion molecule-1 (MAdCAM-1). Furthermore, we generated two kinds of hybridomas producing monoclonal antibodies (mAbs) that recognized Mn 2+ -dependent epitopes of β 7 . Both epitopes were exposed to bind to mAbs on the cells by the introduction of Rap1V12. Although one epitope in the PSI domain of β 7 was exposed on Rap1-deficienct cells, the other epitope in the hybrid domain of β 7 was not. These data indicate that the conversion of Rap1-GDP to GTP exerts two distinct effects stepwise on the conformation of α 4 β 7 . The induction of colitis by Rap1-deficient CD4 + effector/memory T cells suggests that the removal of constraining effect by Rap1-GDP on α 4 β 7 is sufficient for homing of these pathogenic T cells into colon lamina propria (LP).

Background: Lymphocytes circulate between peripheral lymphoid tissues via blood and lymphatic systems, and chemokine-induced migration is important in trafficking lymphocytes to distant sites. The small GTPase Rap1 is important in mediating lymphocyte motility, and Rap1-GEFs are involved in chemokine-mediated Rap1 activation. Here, we describe the roles and mechanisms of Rap1-GEFs in lymphocyte trafficking., Results: In this study, we show that RA-GEF-1 and 2 (also known as Rapgef2 and 6) are key guanine nucleotide exchange factors (GEF) for Rap1 in lymphocyte trafficking. Mice harboring T cell-specific knockouts of Rapgef2/6 demonstrate defective homing and egress of T cells. Sphingosine-1-phosphate (S1P) as well as chemokines activates Rap1 in a RA-GEF-1/2-dependent manner, and their deficiency in T cells impairs Mst1 phosphorylation, cell polarization, and chemotaxis toward S1P gradient. On the other hand, B cell-specific knockouts of Rapgef2/6 impair chemokine-dependent retention of B cells in the bone marrow and passively facilitate egress. Phospholipase D2-dependent production of phosphatidic acid by these chemotactic factors determines spatial distribution of Rap1-GTP subsequent to membrane localization of RA-GEFs and induces the development of front membrane. On the other hand, basal de-phosphorylation of RA-GEFs is necessary for chemotactic factor-dependent increase in GEF activity for Rap1., Conclusions: We demonstrate here that subcellular distribution and activation of RA-GEFs are key factors for a directional movement of lymphocytes and that phosphatidic acid is critical for membrane translocation of RA-GEFs with chemokine stimulation.

Objective: The aim of this study was to evaluate tongue movement and its biomechanical effects during squeezing, one of the oral strategies for processing soft foods, by tongue pressure sensors, videofluorography, and surface electromyography., Design: Fifteen healthy men (mean age, 31.0 ± 4.1 years) without dysphagia were recruited. A 0.1-mm-thick pressure sensor sheet with five measuring points, videofluorography, and surface electromyography were used for synchronous measurements of tongue pressure, hyoid movement, and suprahyoid muscles activity, respectively, while squeezing 5 mL of gels. Amplitude, duration, area, and their sequential order during initial squeezing were analyzed. Differences in hyoid position at the onset, peak, and offset of hyoid movement were also analyzed., Results: At the beginning of initial squeezing, tongue pressure at the middle area of the hard palate, hyoid movement, and suprahyoid muscle activity appeared simultaneously, followed by tongue pressure at the anterior area and then at the posterior area. When the hyoid was in an elevated position, the amplitude of suprahyoid muscle activity and tongue pressure peaked. At the end of initial squeezing, the hyoid position at the offset of hyoid excursion was superior to that at the onset. All evaluation items of tongue pressure, hyoid movement, and suprahyoid muscle activity were modulated according to the texture of gels., Conclusions: During initial squeezing, tongue pressure, hyoid movement, and suprahyoid muscle activity were coordinated while being modulated by the food texture. At the end of initial squeezing, the hyoid was maintained in an elevated position, which might be beneficial for subsequent squeezing., Competing Interests: Declaration of Competing Interest There are no conflict of interests in connection with this article., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The detection of proteases and their complexes with inhibitor proteins is of great importance for diagnosis and medical-treatment applications. In this study, we report a fingerprint-based sensor using an array of single-stranded DNAs (ssDNAs) labeled with environment-responsive 3'-carboxytetramethylrhodamine (TAMRA) for the identification of proteases. Four TAMRA-modified ssDNAs with different sequences solubilized in two different buffer solutions were incorporated in an array that was capable of generating fluorescent fingerprints unique to the proteases through diverse cross-reactive interactions, allowing the discrimination of (i) 8 proteases and (ii) 12 different mixtures of trypsin and its inhibitor protein (α 1 -antitrypsin) by multivariate analysis. Constructing an array with TAMRA-modified DNA aptamers that bind to different sites of human thrombin provides fluorescence fingerprints that reflect a reduction of the exposed surface area of thrombin upon complexation with antithrombin III, even in the presence of human serum. We finally demonstrate the potential of hybridization with complementary DNAs as an effective means to easily double the fingerprint information for proteases. Our approach based on the cross-reactive capability of ssDNAs enables high-throughput fingerprint-based sensing that can be flexibly designed and easily constructed, not only for the identification of a variety of proteins including proteases but also for the evaluation of their complexation ability.

Care food is increasingly required in the advanced-aged society. Mechanical properties of such foods must be modified such that the foods are easily broken by the tongue without chewing. When foods are compressed between the tongue and the hard palate, the tongue deforms considerably, and only soft foods are broken. To simulate tongue compression of soft foods, artificial tongues with stiffness similar to that of the human tongue were created using clear soft materials. Model soft gels were prepared using gellan gums. A piece of gel on an artificial tongue was compressed using a texture analyzer. The deformation profile during the compression test was obtained using a video capture system. The soft machine equipped a soft artificial tongue sometimes fractured food gels unlike hard machine, which always fracture gels. The fracture properties measured using the soft machine were better than those obtained from a conventional test between hard plates to mimic natural oral processing in humans. The fracture force on foods measured using this soft machine may prove useful for the evaluation of food texture that can be mashed using the tongue., Competing Interests: The authors declare no conflict of interest.

A one-component array-based sensor consisting of a dansyl-modified polylysine (PLL-Dnc) is capable of discriminating the types and compositional ratios of human cells using varying buffer conditions. The PLL-Dnc sensor array, which affords turn-on fluorescence responses against analyte cells that depend on the pH value and the ionic strength, generates differential fluorescence signatures of cells and successfully discriminates eight types of human cell lines (2.0 × 10 4 cells/mL) with 100% accuracy using multivariate analysis. The array also allows differentiation of the composition of the cell mixtures that contain cells with the same tissue origin but of different subtypes. The good discrimination ability and simple platform of our "one-component"-based array allows an easy and rapid sensing of cells without requiring any information on specific biomarkers.

Several bacterial moonlighting proteins act as adhesion factors, which are important for bacterial colonization of the gastrointestinal (GI) tract. However, little is known about the adherence properties of moonlighting proteins in the GI tract. Here, we describe a new approach for visualizing the localization of moonlighting protein-coated fluorescent microbeads in the whole GI tract by using a tissue optical clearing method, using elongation factor Tu (EF-Tu) as an example. As a bacterial cell surface-localized protein mimic, recombinant EF-Tu from Lactobacillus reuteri was immobilized on microbeads. EF-Tu-coating promoted the interaction of the microbeads with a Caco-2 cell monolayer. Next, the microbeads were orally administered to mice. GI whole tissues were cleared in aqueous fructose solutions of increasing concentrations. At 1 h after administration, the microbeads were diffused from the stomach up to the cecum, and after 3 h, they were diffused throughout the intestinal tract. In the lower digestive tract, EF-Tu-beads were significantly more abundant than non-coated control beads, suggesting that EF-Tu plays an important role in the persistence of the microbeads in the GI tract. The new approach will help in evaluating how moonlighting proteins mediate bacterial colonization.

Mimicking sensory principles encountered in animals, whereby numerous tastants and odorants are identified based on "pattern"-like sensory inputs that are generated by arrays of sensory cells, allows creating a unique technique that is distinct from conventional chemical sensing systems, as the latter usually require specific recognition of target analytes. Herein, we present a highly discriminative small fluorescent array of block copolymers that can recognize various bioanalytes in a biomimicry manner. These polyethylene glycol/poly-l-lysine block copolymers are functionalized with fluorescein as a fluorescent reporter unit and hydrophobic amino acids as cross-reactive recognition units, which provides the ability to generate fluorescent response patterns unique to proteins and cells. Multivariate analysis on the patterns obtained with an array consisting of solely 3 block copolymers allowed identifying not only 20 proteins and 10 mammalian cells individually but also complex protein mixtures with slightly different compositions. This design guideline for creating a versatile biomimicry sensing system, which is based on the bifunctionalization of polymeric materials, is expected to offer a powerful platform for simple and high-throughput sensing of a wide variety of bioanalytes.

Introduction: Collagen peptides have been widely used as a food supplement. After ingestion of collagen peptides, oligopeptides containing hydroxyproline (Hyp), which are known to have some physiological activities, are detected in peripheral blood. However, the effects of collagen-peptide administration on immune response are unclear. In the present study, we tested the effects of collagen-peptide ingestion on allergic response and the effects of collagen-derived oligopeptides on CD4 + T-cell differentiation., Methods: BALB/c mice fed a collagen-peptide diet were immunized with ovalbumin (OVA), and their serum IgE and IgG levels, active cutaneous anaphylaxis, and cytokine secretion by splenocytes were examined. Naive CD4 + T cells were stimulated with anti-CD3 and anti-CD28 in the presence of collagen-derived oligopeptides, and the expression of IFN-γ, IL-4, and Foxp3 was analyzed., Results: In an active anaphylaxis model, oral administration of collagen peptides suppressed serum OVA-specific immunoglobulin E (IgE) production and diminished anaphylaxis responses. In this model, the ingestion of collagen peptides skewed the pattern of cytokine production by splenocytes toward T-helper (Th) type 1 and regulatory T (Treg) cells. In vitro T-helper cell differentiation assays showed that Hyp-containing oligopeptides promoted Th1 differentiation by upregulating IFN-γ-induced signal transducer and activator of transcription 1 (STAT1) signaling. These oligopeptides also promoted the development of Foxp3 + Treg cells in response to antigen stimulation in the presence of TGF-β., Conclusions: Collagen-peptide ingestion suppresses allergic responses by skewing the balance of CD4 + T cells toward Th1 and Treg cells and seems to be a promising agent for preventing allergies and inflammatory diseases., (© 2018 The Authors. Immunity, Inflammation and Disease Published by John Wiley & Sons Ltd.)

Signature-based protein sensing has recently emerged as a promising prospective alternative to conventional lock-and-key methods. However, most of the current examples require the measurement of optical signals from spatially-separated materials for the generation of signatures. Herein, we present a new approach for the construction of multi-fluorescent sensing systems with high accessibility and tunability, which allows generating protein fluorescent signatures from a single microplate well. This approach is based on conjugates between nano-graphene oxide (nGO) and three single-stranded DNAs (ssDNAs) that exhibit different sequences and fluorophores. Initially, the three fluorophore-modified ssDNAs were quenched simultaneously by binding to nGO. Subsequent addition of analyte proteins caused a partial recovery in fluorescent intensity of the individual ssDNAs. Based on this scheme, we have succeeded in acquiring fluorescence signatures unique to (i) ten proteins that differ with respect to pI and molecular weight and (ii) biochemical marker proteins in the presence of interferent human serum. Pattern-recognition methods demonstrated high levels of discrimination for this system. The high discriminatory power and simple format of this sensor system should enable an easy and fast evaluation of proteins and protein mixtures., Competing Interests: The authors declare no conflict of interest.

The identification of post-translational modifications (PTMs) in proteins has been of particular interest in the elucidation of human diseases and the improvement of therapeutic proteins. Herein, we report a novel strategy toward the construction of fingerprint-based PTM-sensing systems as an alternative to conventional specific recognition tools. Our strategy is based on poly-l-lysine (PLL) derivatives with two distinct properties suitable to fingerprint-based protein-sensing: (i) a turn-on fluorescent signal upon binding to proteins and (ii) condition-dependent cross-reactivity toward proteins and PTMs. One type of PLL derivative under varying solution properties (pH value and ionic strength) was sufficient to construct a sensing array that produces unique fluorescence fingerprints for structurally similar mammalian albumins with/without a wide variety of chemical modifications corresponding to PTMs. This approach was also applicable for the recognition of deviations in physicochemical properties of proteins as a result of realistic glycation and phosphorylation events. Multivariate analyses of the thus obtained fingerprints successfully identified analytes with 100% accuracy (qualitatively and quantitatively) in all cases. This study thus demonstrates for the first time a fingerprint-based sensing of proteins with/without PTMs using a single, highly accessible, and tunable synthetic polymer, and accordingly offers a powerful platform for simple high-throughput sensing of PTMs in proteins.

The objectives of this study were to examine the effects of mouthful quantities and mechanical properties of gels on natural mastication behaviors using electromyography (EMG). Two types of hydrocolloid gels (A and K) with similar fracture loads but different moduli and fracture strains were served to eleven normal women in 3-, 6-, 12-, and 24-g masses in a randomized order. EMG activities from both masseter muscles were recorded during natural mastication. Because of the similar fracture loads, the numbers of chews, total muscle activities, and entire oral processing times were similar for similar masses of both gel types. Prior to the first swallow, the more elastic K gel with a higher fracture strain required higher muscle activities than the brittle A gel, which had higher modulus. Majority of subjects had preferred sides of chewing, but all subjects with or without preferred sides used both masseters during the consumption of gels. Similar effects of masses and types of gels were observed in EMG activities of both sides of masseters. Contributions of the dominant side of chewing were diminished with increasing masses of gels, and the mass dependency on ratio of the dominant side was more pronounced with K gel. More repetitions of smaller masses required greater muscle activities and longer periods for the consumption of 24-g gel portions. Reduction in the masses with an increased number of repetitions necessitated slower eating and more mastication to consume the gel portions. These observations suggest that chewing using both sides is more effective and unconsciously reduces mastication times during the consumption of gels., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Rap1-GTP activates leukocyte function-associated antigen-1 (LFA-1) to induce arrest on the high endothelial venule (HEV). Here we show that Rap1-GDP restrains rolling behaviours of T cells on the peripheral lymph node addressin (PNAd), P-selectin and mucosal addressin cell adhesion molecule-1 (MadCAM-1) by inhibiting tether formation. Consequently, Rap1 deficiency impairs homing of naive T cells to peripheral lymph nodes, but accelerates homing of TH17 and TH1 cells to the colon, resulting in spontaneous colitis with tumours. Rap1-GDP associates with and activates lymphocyte-oriented kinase, which phosphorylates ERM (ezrin, radixin and moesin) in resting T cells. Phosphomimetic ezrin reduces the rolling of Rap1-deficient cells, and thereby decreases their homing into the colon. On the other hand, chemokines activate Rap1 at the plasma membrane within seconds, and Rap1-GTP binds to filamins, which diminishes its association with the β2 chain of LFA-1 and results in LFA-1 activation. This Rap1-dependent regulation of T-cell circulation prevents the onset of colitis.

In lymphocytes, the kinase Mst1 is required for the proper organization of integrins in the plasma membrane at the leading edge of migrating cells, which is critical for lymphocyte trafficking. We found a functional link between the small G protein Rab13 and Mst1 in lymphocyte adhesion and migration. In response to stimulation of T lymphocytes with chemokine, Mst1 promoted phosphorylation of the guanine nucleotide exchange factor DENND1C (differentially expressed in normal and neoplastic cells domain 1C), which activated Rab13. Active Rab13 associated with Mst1 to facilitate the delivery of the integrin LFA-1 (lymphocyte function-associated antigen 1) to the leading edge of lymphocytes. Delivery of LFA-1 involved the recruitment of myosin Va along actin filaments, which extended as a result of the localization of the actin regulatory protein VASP to the cell periphery through phosphorylation of VASP at Ser(157) by Mst1. Inhibition of Rab13 function reduced the adhesion and migration of lymphocytes on ICAM-1 (intercellular adhesion molecule-1), the ligand for LFA-1, and inhibited the formation of a ring-like arrangement of LFA-1 at the contact sites between T cells and antigen-presenting cells. The lymphoid tissues of Rab13-deficient mice had reduced numbers of lymphocytes because of the defective trafficking capability of these cells. These results suggest that Rab13 acts with Mst1 to regulate the spatial distribution of LFA-1 and the motility and trafficking of lymphocytes., (Copyright © 2014, American Association for the Advancement of Science.)

Background: In the recent hyper-aged societies of developed countries, the market for soft diets for patients with dysphagia has been growing and numerous jelly-type foods have become available. However, interrelationships between the biomechanics of oral strategies and jelly texture remain unclear. The present study investigated the influence of the initial consistency of jelly on tongue motor kinetics in different oral strategies by measuring tongue pressure against the hard palate., Methods: Jellies created as a mixture of deacylated gellan gum and psyllium seed gum with different initial consistencies (hard, medium or soft) were prepared as test foods. Tongue pressure production while ingesting 5 ml of jelly using different oral strategies (Squeezing or Mastication) was recorded in eight healthy volunteers using an ultra-thin sensor sheet system. Maximal magnitude, duration and total integrated values (tongue work) of tongue pressure for size reduction and swallowing in each strategy were compared among initial consistencies of jelly, and between Squeezing and Mastication., Results: In Squeezing, the tongue performed more work for size reduction with increasing initial consistency of jelly by modulating both the magnitude and duration of tongue pressure over a wide area of hard palate, but tongue work for swallowing increased at the posterior-median and circumferential parts by modulating only the magnitude of tongue pressure. Conversely, in Mastication, the tongue performed more work for size reduction with increasing initial consistency of jelly by modulating both magnitude and duration of tongue pressure mainly at the posterior part of the hard palate, but tongue work as well as other tongue pressure parameters for swallowing showed no differences by type of jelly., Conclusions: These results reveal fine modulations in tongue-palate contact according to the initial consistency of jelly and oral strategies.

Proanthocyanidins, also known as condensed tannins and/or oligomeric flavonoids, occur in many edible plants and have various interesting biological activities. Previously, we reported a synthetic method for the preparation of various procyanidins in pure form and described their biological activities. Here, we describe the synthesis of procyanidin B1 acetylated analogs and discuss their inhibition activities against HeLa S3 cell proliferation. Surprisingly, the lower-unit acetylated procyanidin B1 strongly inhibited the proliferation of HeLa S3 cells. This molecule showed much stronger inhibitory activity than did epigallocatechin-3-O-gallate (EGCG), green tea polyphenol, and dimeric compounds that included EGCG as a unit. This result suggests that the phenolic hydroxyl groups of the upper-units in flavan-3-ols are important for their inhibitory activity against cancer cell proliferation and that a hydrophobic lower unit dimer enhances this activity.

This paper proposes a novel texture sensing method for nursing-care gel by using an artificial mastication system, in which not only mechanical characteristics but also geometrical ones are objectively and quantitatively evaluated. When human masticates gel food, she or he perceives the changes of the shape and contact force simultaneously. Based on the impressions, they evaluate the texture. For reproducing such a procedure, the pressure distribution of gel is measured in the simple artificial mastication, and the information associated to both the geometrical and mechanical characteristics is simultaneously acquired. The relationship between the value of sensory evaluation (i.e. impression human perceives), and the pressure distribution data is numerically modeled by applying the image texture analysis. Experimental results show that the proposed method succeeds in estimating the values of sensory evaluation of nine kinds of gel with the coefficient of determination greater than 0.93.

Semisynthetic cephalosporins, the best-selling antibiotics worldwide, are derived from 7-aminocephalosporanic acid (7-ACA). Currently, in the pharmaceutical industrie, 7-ACA is mainly produced from cephalosporin C by sequential application of D-amino acid oxidase and cephalosporin acylase. Here we study the potential of industrially amenable enzyme gamma-glutamyltranspeptidase from Bacillus subtilis for 7-ACA production, since the wild-type gamma-glutamyltranspeptidase of B. subtilis has inherent glutaryl-7-aminocephalosporanic acid acylase activity with a k(cat) value of 0.0485 s(-1). Its activity has been enhanced by site directed and random mutagenesis. The k(cat)/K(m) value was increased to 3.41 s(-1) mM(-1) for a E423Y/E442Q/D445N mutant enzyme and the kcat value was increased to 0.508 s(-1) for a D445G mutant enzyme. Consequently, the catalytic efficiency and the turnover rate were improved up to about 1000-fold and 10-fold, compared with the wildtype gamma-glutamyltranspeptidase of B. subtilis.

The textural properties of cooked rice were investigated in the presence and the absence of gum arabic (GA) and soybean soluble polysaccharide (SSPS). SSPS was more effective in increasing the hardness and in decreasing the stickiness of the rice grains than GA. For both polysaccharides, the increase in hardness was more apparent in the whole body than at the periphery, whereas the decrease in stickiness was more apparent at the periphery than in the whole body. SSPS was more effective in retarding the gelatinization of rice starch and in lowering the elastic characters of the glutinous layer (the materials leached out of the rice grains during cooking) along with a decrease in the amount of amylopectin leached. The textural hardness of cooked rice was determined by the degree of starch gelatinization, whereas the textural stickiness was related to the rheological characters of the glutinous layer and the leaching profile of the starch components.

Aqueous solutions of gellan gum with comparable molecular masses but with different acyl contents were investigated by atomic force microscopy and rheological measurements in the presence or absence of various cations. For a high-acyl sample, no continuous network structures were identified microscopically, except in the presence of Ca (2+), where structural inhomogeneity was the highest in terms of the height distribution of molecular assemblies. Rheological thermal hysteresis between sol-gel transitions was detected in the presence of K (+) and Ca (2+), particularly Ca (2+). The storage modulus at 20 degrees C was larger in the order Na (+) < Ca (2+) < K (+). For a low-acyl sample, continuous network structures were identified in the presence of each cation, involving greater thermal hysteresis than the corresponding data for the high-acyl sample. Structural homogeneity was the highest in the presence of K (+). Thermal hysteresis and elasticity of the system were discussed in terms of continuousness and homogeneity of network structures.

7-Aminocephalosporanic acid (7-ACA) is an important material in the production of semisynthetic cephalosporins, which are the best-selling antibiotics worldwide. 7-ACA is produced from cephalosporin C via glutaryl-7-ACA (GL-7-ACA) by a bioconversion process using d-amino acid oxidase and cephalosporin acylase (or GL-7-ACA acylase). Previous studies demonstrated that a single amino acid substitution, D433N, provided GL-7-ACA acylase activity for gamma-glutamyltranspeptidase (GGT) of Escherichia coli K-12. In this study, based on its three-dimensional structure, residues involved in substrate recognition of E. coli GGT were rationally mutagenized, and effective mutations were then combined. A novel screening method, activity staining followed by a GL-7-ACA acylase assay with whole cells, was developed, and it enabled us to obtain mutant enzymes with enhanced GL-7-ACA acylase activity. The best mutant enzyme for catalytic efficiency, with a k(cat)/K(m) value for GL-7-ACA almost 50-fold higher than that of the D433N enzyme, has three amino acid substitutions: D433N, Y444A, and G484A. We also suggest that GGT from Bacillus subtilis 168 can be another source of GL-7-ACA acylase for industrial applications.

The aspartyl residue at position 433 of gamma-glutamyltranspeptidase of Escherichia coli K-12 was replaced by an asparaginyl residue. This substitution enabled gamma-glutamyltranspeptidase to deacylate glutaryl-7-aminocephalosporanic acid, producing 7-aminocephalosporanic acid, which is a starting material for the synthesis of semisynthetic cephalosporins.

An enzymatic method for synthesizing various gamma-D-glutamyl compounds efficiently and stereospecifically involving bacterial gamma-glutamyltranspeptidase (EC 2.3.2.2) with D-glutamine as a gamma-glutamyl donor was developed. With D-glutamine as a gamma-glutamyl donor instead of L-glutamine in gamma-glutamyltaurine synthesis, by-products such as gamma-glutamylglutamine and gamma-glutamyl-gamma-glutamyltaurine were not synthesized and the yield of gamma-glutamyltaurine dramatically increased from 25 to 71%. It was also shown that the purification could be simplified without these gamma-glutamyl by-products. The possibility of synthesizing various gamma-D-glutamyl compounds was also shown.