Investigation of the relevance between cell cycle status and the bioactivity of exogenously delivered biomacromolecules is hindered by their time-consuming cell internalization and the cytotoxicity ...of transfection methods. In this study, we addressed these problems by utilizing the photochemical internalization (PCI) method using a peptide/protein-photosensitizer conjugate, which enables immediate cytoplasmic internalization of the bioactive peptides/proteins in a light-dependent manner with low cytotoxicity. To identify the cell-cycle dependent apoptosis, a TatBim peptide-photosensitizer conjugate (TatBim-PS) with apoptotic activity was photo-dependently internalized into HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (Fucci2). Upon irradiation, cytoplasmic TatBim-PS internalization exceeded 95% for all cells classified in the G
, S, and G
/M cell cycle phases with no significant differences between groups. TatBim-PS-mediated apoptosis was more efficiently triggered by photoirradiation in the G
/S transition than in the G
and S/G
/M phases, suggesting high sensitivity of the former phase to Bim-induced apoptosis. Thus, the cell cycle dependence of Bim peptide-induced apoptosis was successfully investigated using Fucci2 indicator and the PCI method. Since PCI-mediated cytoplasmic internalization of peptides is rapid and does not span multiple cell cycle phases, the Fucci-PCI method constitutes a promising tool for analyzing the cell cycle dependence of peptides/protein functions.
Background
Ulcerative colitis (UC) is a chronic inflammatory disease of the colon with an intractable, recurrent course. The goal of UC therapy is to target mucosal healing because immune-suppressive ...therapy for UC frequently results in relapse. However, few drugs directly target mucosal healing. We, therefore, aim to evaluate the therapeutic effect of an investigational drug on intestinal epithelial cells in an in vitro UC model using human colonic organoids.
Methods
Colonic organoids were isolated from human colon and cultured. A mixture of cytokines and bacterial components were used to mimic UC in humans. The effect of the investigational drug on colonic organoid was evaluated by microarray analysis and 3D immunofluorescence. The enrichment of stem cells was assessed with a colony formation assay.
Results
Inflammatory stimulation resulted in a significant induction of inflammatory-related genes in colonic organoids whereas cell differentiation was suppressed. Treatment with the investigational drug KAG-308 showed reciprocal dynamics of gene expression to inflammatory stimulation, which resulted in not only the suppression of immune response but also the promotion of cellular differentiation towards secretory lineages. Moreover, SPDEF and Reg4 were identified as novel targets for the enrichment of intestinal epithelial stem cells and mucosal healing.
Conclusions
The establishment of in vitro UC model using human colonic organoid could reveal the effects and targets of investigational drugs in intestinal epithelial cells under inflammation conditions. Further maturation of this system might be more efficient to predict the effect on UC, as compared with the use of animal model, for the development of new drugs.
Quasicrystalline structures and aperiodic metamaterials find applications ranging from established consumer gadgets to potential high‐tech photonic components owing to both complex arrangements of ...constituents and exotic rotational symmetries. Magnonics is an evolving branch of magnetism research where information is transported via magnetization oscillations (magnons). Their control and manipulation are so far best accomplished in periodic metamaterials which exhibit properties artificially modulated on the nanoscale. They give rise to functional components, such as band stop filters, magnonic transistors and nanograting couplers. Here, spin‐wave excitations in artificial ferromagnetic quasicrystals created via aperiodic arrangement of nanoholes are studied experimentally. Their ten‐fold rotational symmetry results in multiplexed magnonic nanochannels, suggesting a width down to 50 nm inside a so‐called Conway worm. Key elements of design are emergent magnon motifs and the worm‐like features which are scale‐invariant and not present in the periodic metamaterials. By imaging wavefronts in quasicrystals, insight is gained into how the discovered features materialize as a dense wavelength division multiplexer.
Artificial ferromagnetic quasicrystals (AMQs) allow spin waves to form nanochannels, which incorporate peculiar sequences of bends, and thus the wavelength of the spin waves is different from nanochannel to nanochannel at a single input frequency. The results suggest that AMQs promise a new class of magnonic devices such as ultra‐compact and dense wavelength division multiplexers.
Synovial sarcoma (SS) is an aggressive tumor that most often affects the deep soft tissues in young adults. Intrathoracic SS is rare and is associated with poor outcome, highlighting the urgent need ...for a novel therapeutic strategy. In the process of clinical sequencing, we identified two patients with intrathoracic SS harboring the BRAF V600E mutation. The patients were women aged 32 and 23 years, and both presented with SS18–SSX2-positive monophasic SS in the thoracic cavity. BRAF V600E mutations were detected by next generation sequencing, and validated immunohistochemically by diffuse intense positivity to BRAF V600E mutation-specific antibodies. The phosphorylated ERK (pERK) immunohistochemistry result was also positive. One patient received a combination therapy of dabrafenib and trametinib, which led to tumor shrinkage. However, the tumor growth progressed 7.5 months later with an additional NRAS Q61K mutation. Immunohistochemical screening of 67 archival SS tumor samples failed to identify additional samples with BRAF V600E mutation. However, 32% of BRAF V600E-negative cases was positive for pERK, and one of the six tumors showing the highest pERK expression harbored an FGFR2-activating mutation. This is the first report of targetable BRAF mutation in a small subset of SS. Our study suggests involvement of the mitogen-activated protein kinase pathway and the potential clinical implication of BRAF mutation screening in SS.
The coral microbiome has attracted increased attention because of its potential roles in host protection against deadly diseases. However, little is known about the role of coral-associated bacteria ...against the temperature-dependent opportunistic pathogen
Vibrio coralliilyticus
. In this study, we tested whether bacteria associated with the reef-building coral
Galaxea fascicularis
could inhibit the growth of
V. coralliilyticus
. Twenty-nine cultivable bacteria were successfully isolated from a healthy colony of
G. fascicularis
kept in an aquarium. Among the bacterial isolates, three
Ruegeria
sp. strains inhibited the growth of
V. coralliilyticus
P1 as a reference strain and
Vibrio
sp. isolated in this study.
Ruegeria
sp. strains were also detected from other
G. fascicularis
colonies in the aquarium and in previous field studies by 16S rRNA amplicon sequencing, suggesting that
Ruegeria
sp. strains are common among
G. fascicularis
colonies. These results illuminate the potential role of
Ruegeria
sp. in protecting corals against pathogenic
Vibrio
species.
Rechargeable batteries using bis‐ or tris‐fused tetrathiafulvalenes as an active electrode material exhibit good cycle performance by controlling the number of electrons participating in the redox ...reaction. The TTPY cell using a four‐electron redox process shows a discharge capacity of 168 mAh g−1 and good cycle‐life stability of 84 % of the initial capacity after 100 cycles (see charge‐discharge curves).
We aimed to clarify the communication behaviors between trainee dentists and simulated patients (SPs), to examine how the level of trainee dentists' self-reported empathy influences assessment by SPs ...in medical interviews.
The study involved 100 trainee dentists at Okayama University Hospital and eight SPs. The trainee dentists conducted initial interviews with the SPs after completing the Japanese version of the Jefferson Scale of Empathy (JSE). All interviews were recorded and analyzed using the Roter Interaction Analysis System (RIAS). The SPs assessed the trainees' communication immediately after each interview. The trainee dentists were classified into two groups (more positive and less positive) according to SP assessment scores.
Compared with less-positive trainees, the more-positive trainees scored higher in the RIAS category of emotional expression and lower in the medical data gathering category. There was no difference in dental data gathering between the two groups. SP ratings for more-positive trainees were higher for use of positive talk and emotional expression and lower for giving medical information and dental information. Trainees with more positive ratings from SPs had significantly higher JSE total scores.
The results of this study suggest that responding to the emotions of patients is an important behavior in dentist-patient communication, according to SPs' positive assessment in medical interviews. Further, SPs' assessment of trainees' communication was related to trainees' self-reported empathy, which indicates that an empathic attitude among dentists is a significant determinant of patient satisfaction.
Adults with food-protein-induced enterocolitis syndrome (FPIES) often develop severe abdominal symptoms after eating seafood. However, no investigation of a food elimination strategy for adult FPIES ...patients has been performed to date.
We conducted a retrospective cohort study of seafood–avoidant adults by telephone interview, based on the diagnostic criteria for adult FPIES reported by González et al. We compared the clinical profiles, abdominal symptoms, and causative seafoods between FPIES and immediate-type food allergy (IgE-mediated FA) patients. We also profiled the detailed intake-status of seafoods in adult FPIES patients.
Twenty-two (18.8 %) of 117 adults with seafood-allergy were diagnosed with FPIES. Compared with the IgE-mediated FA patients, FPIES patients had an older age of onset, more pre-existing gastrointestinal and atopic diseases, more episodes, longer latency and duration of symptoms, more nausea, abdominal distention, and severe abdominal pain, and more frequent vomiting and diarrhea. In particular, abdominal distention—reflecting intestinal edema and luminal fluid retention—may be the most distinctive characteristic symptom in adult FPIES (p < 0.001). Bivalves, especially oysters, were the most common cause of FPIES. Strikingly, intake-status profiling revealed that many FPIES patients can safely ingest an average of 92.6 % of seafood species other than the causative species.
There are many differentiators between FPIES and IgE-mediated FA, which may reflect differences in the underlying immunological mechanisms. Although seafood FPIES is unlikely to induce tolerance, many patients can ingest a wide variety of seafood species after a long period from onset.
Tumor protein p53 (TP53) mutation is a well-known occurrence at the late phase of carcinogenesis during the adenoma-carcinoma sequence of a sporadic colon cancer. Although numerous reports about ...clinical information of the patients with colon cancer have suggested that
mutation might be related to various types of malignant potential, the direct effects of this mutation on the malignant potential of colon cancer remain unknown. Notably, no previous report has described a relationship between
mutation and cancer stemness. We therefore aimed to assess the function of a TP53 mutant induced by the CRISPR-Cas9 system in colon cancer cells. In this study, two
mutations, corresponding to exon 3 (TP53E3) and 10 (TP53E10), were generated in LS174T cells derived from a wild-type TP53 human colon cancer via a lentiviral CRISPR-Cas9 system. The loss of function of TP53 resulting from both mutations manifested as resistance to Nutlin3a-induced apoptosis and the downregulation of target genes of TP53. TP53 mutants exhibited an enhanced malignant potential, characterized by accelerated cell growth, invasiveness, chemoresistance, and cancer stemness. Interestingly, TP53E10 but not TP53E3 cells exhibited aberrant transcriptional activity of regenerating family member 1-α (
) and expression of REG1A, resulting in the acquisition of enhanced malignant potential. In conclusion, we demonstrated for the first time that
genomic mutation into human colon cancer cells affects the malignant potential. IMPLICATIONS: These findings suggest that both a loss of function and an aberrant gain of function of TP53 might promote high malignant potentials at the late phase of carcinogenesis in colon cancer.
We developed a new perovskite film forming process based on solvent bathing method. The preheating of substrate before spin-coating of precursor solution drastically changed the quality of the ...obtained perovskite films. The perovskite films formed by modified solvent bathing method were much dense with large grains. The improvement of open circuit voltage implied that the modification succeeded to form the better interface between TiO2 and perovskite layer and to grow the higher quality perovskite layer. Finally, we obtained the best conversion efficiency of 12.6%. Although the conversion efficiency of this work is not sufficient compared with other reports, our modified solvent bathing method has a wider process window comparing to the commonly used anti-solvent dipping methods. We expect that our modified method can also applied to monolithic tandem solar cells.
•The solvent bathing method with preheating of substrate was developed.•Obtained perovskite films were much dense with large grains.•The improvement of open circuit voltage due to a better interface between TiO2 and perovskite layers.•The modified solvent bathing method has a wider process window.
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