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The accurate thermosensing requires a minimum impact of autofluorescence and light scattering from the samples. In this study, we discovered that commercially available benzene-1,4-diboronic acid (BDBA) doped co-crystals with trimethylolpropane (TMP) exhibit excellent thermochromic dual phosphorescence properties over a wide temperature range from -132 to 40 °C, despite its simple structure that does not have any donor-acceptor linkage. The dual phosphorescence was consisted of monomeric benzene-1,4-diboronate (BDBA ester) and aggregation-stabilized species. With an increase in temperature, the emission intensity from the monomeric state significantly decreased, whereas that originating from the aggregated state remained almost constant owing to the difference in their thermal stabilities. Further investigation revealed that molecular distortions in singlet excited states enable efficient intersystem crossing, causing efficient phosphorescence from the monomeric state of BDBA ester.
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Boronate assemblies, derived from benzene-1,4-diboronic acid, showed room-temperature phosphorescence (RTP) in both the solid-state and dispersions, being quite sensitive to the preparation conditions. Our chemometrics-assisted quantitative structure-property relationship (QSPR) analysis between the nanostructure and RTP behavior of boronate assemblies led us not only to understand their RTP mechanism but also to predict the RTP properties of unknown assemblies from the PXRD patterns.
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Bacillus subtilis natto is used in the production of natto, a traditional fermented soy food, and has beneficial immunomodulatory effects in humans. Single-stranded RNA (ssRNA) viruses, including influenza and coronavirus, often cause global pandemics. We proposed a human cell culture model mimicking ssRNA viral infection and investigated the ability of B. subtilis natto to induce antiviral effects in the model. The gene expressions were analyzed using quantitative real-time reverse transcription PCR. M1-phenotype macrophages derived from THP-1 cells strongly express the Toll-like receptor 8 (76.2-hold), CD80 (64.2-hold), and CCR7 (45.7-hold) mRNA compared to M0 macrophages. One µg/mL of resiquimod (RSQ)-stimulation induced the expression of IRF3 (1.9-hold), CXCL10 (14.5-hold), IFNß1 (3.5-hold), ISG20 (4.4-hold), and MxA (1.7-hold) mRNA in the M1-phenotype macrophages. Based on these results, the RSQ-stimulated M1-phenotype macrophages were used as a cell culture model mimicking ssRNA viral infection. Moreover, the B. subtilis natto XF36 strain induced the expression of genes associated with antiviral activities (IFNß1, IFNλ1, ISG20, and RNase L) and anti-inflammatory activities (IL-10) in the cell culture model. Thus, it is suggested that the XF36 suppresses viral infections and excessive inflammation by inducing the expression of genes involved in antiviral and anti-inflammatory activities.
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An afterglow is beneficial as an emission signal in the field of displays and imaging probes. Here, boronic acid-appended and spirolactam ring-containing rhodamine dye 1 was synthesized and grafted onto the surface of room-temperature phosphorescence-active boronate nanoparticles (BPs), composed of polymeric 3-benzo-2,4,8,10-tetraoxa-3,9-diboraspiro[5.5]undecane. The resultant ensemble, 1@BP, exhibited a greenish afterglow. However, the addition of Al3+ into the dispersion solution with 1@BP led to a change in the afterglow to grass green as a result of Förster-type energy transfer from the phosphorescent BP to the Al3+ -interacting rhodamine dye 1 on the surface. Based on the ratio of the two emission intensities, a linear response in the concentration range of 3.8-15.2â µM was observed, with a detection limit of 4.2â µM for Al3+ . A metal ion-dependent discernable color in afterglow was observed on a 1@BP-coated filter paper, which would be useful for not only film-based afterglow chemosensors but also encryption application.
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Aluminio , Agua , Iones , Metanol , RodaminasRESUMEN
To implement precision oncology, analytical validity as well as clinical validity and utility are important. However, proficiency testing (PT) to assess validity has not yet been systematically performed in Japan. To investigate the quality of next-generation sequencing (NGS) platforms and cancer genome testing prevalent in laboratories, we performed pilot PT using patient samples. We prepared genomic DNA from the cancer tissue and peripheral blood of 5 cancer patients and distributed these to 15 laboratories. Most participating laboratories successfully identified the pathogenic variants, except for two closely located KRAS variants and 25 bp delins in EGFR. Conversely, the EGFR L858R variant was successfully identified, and the allele frequency was similar for all the laboratories. A high DNA integrity number led to excellent depth and reliable NGS results. By conducting this pilot study using patient samples, we were able to obtain a glimpse of the current status of cancer genome testing at participating laboratories. To enhance domestic cancer genome testing, it is important to conduct local PT and to involve the parties concerned as organizers and participants.
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NeoplasiasRESUMEN
Two types of boronic acid-appended D-π-A cyanostilbenes were synthesized to produce chiral boronate ensembles via dehydration with tartaric acid. The aggregation-induced high sensitivity and positional effect of the CN group on the emission properties allowed for chemometrics-coupled chiral recognition.
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Natto is a traditional Japanese fermented soy product high in γ-polyglutamic acid (γ-PGA), whose beneficial effects have been reported. We prepared high-γ-PGA natto and compared the dietary influence on liver lipids and cecal microbiota in mice fed a diet containing it or a standard diet. The mice were served a 30% high-γ-PGA natto diet (PGA group) or standard diet (Con group) for 28 days. Liver lipids, fecal lipids, and fecal bile acids were quantified. Cecal microbiota were analyzed by PCR amplification of the V3 and V4 regions of 16S rRNA genes and sequenced using a MiSeq System. Additionally, the cecal short-chain fatty acid profile was assessed. The results revealed that the liver lipid and triglyceride contents were significantly lower (p<0.01) and amounts of bile acids and lipids in the feces were significantly higher in the PGA group than in the Con group. The cecal butyric acid concentration was observed to be significantly higher in the PGA group than in the Con group. Principal component analysis of the cecal microbiota revealed that the PGA and Con groups were distinct. The ratio of Firmicutes/Bacteroidetes was found to be significantly low in the PGA mice. The results revealed a significantly higher relative abundance of Lachnospiraceae (p<0.05) and significantly lower relative abundance of Coriobacteriaceae (p<0.01) in the PGA group. Analysis of the correlation between bacterial abundance and liver lipids, cecal short-chain fatty acids, fecal lipids, and fecal bile acids suggested that intestinal microbiota can be categorized into different types based on lipid metabolism. Hepatic lipid accumulation typically facilitates the onset of nonalcoholic fatty liver disease (NAFLD). Our findings suggest that high-γ-PGA natto is a beneficial dietary component for the prevention of NAFLD.
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The newly prepared fluorescent carboxyamidoquinolines (1-3) and their Zn(II) complexes (Zn@1-Zn@3) were used to bind and sense various phosphate anions utilizing a relay mechanism, in which the Zn(II) ion migrates from the Zn@1-Zn@3 complexes to the phosphate, namely adenosine 5'-triphosphate (ATP) and pyrophosphate (PPi), a process accompanied by a dramatic change in fluorescence. Zn@1-Zn@3 assemblies interact with adenine nucleotide phosphates while displaying an analyte-specific response. This process was investigated using UV-vis, fluorescence, and NMR spectroscopy. It is shown that the different binding selectivity and the corresponding fluorescence response enable differentiation of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), pyrophosphate (PPi), and phosphate (Pi). The cross-reactive nature of the carboxyamidoquinolines-Zn(II) sensors in conjunction with linear discriminant analysis (LDA) was utilized in a simple fluorescence chemosensor array that allows for the identification of ATP, ADP, PPi, and Pi from 8 other anions including adenosine 5'-monophosphate (AMP) with 100 % correct classification. Furthermore, the support vector machine algorithm, a machine learning method, allowed for highly accurate quantitation of ATP in the range of 5-100â µM concentration in unknown samples with error <2.5 %.
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Adenosina Trifosfato , Zinc , Adenosina Difosfato , Adenosina Monofosfato , Fluorescencia , Espectroscopía de Resonancia Magnética , Espectrometría de FluorescenciaRESUMEN
We found that boronate particles (BP), as a self-assembled system prepared by sequential dehydration of benzene-1,4-diboronic acid with pentaerythritol, showed greenish room-temperature phosphorescence (RTP). This emission was observed in both solid and dispersion state in water. To understand the RTP properties, X-ray crystallographic analysis, and density functional theory (DFT) and time-dependent DFT at M06-2X/6-31G(d,p) level were performed using 3,9-dibenzo-2,4,8,10-tetraoxa-3,9-diboraspiro[5.5]undecane (1) as a model compound. Our interest in functionalizing the RTP-active particles led us to graft Rhodamine B onto their surface. The resulting system emitted a dual afterglow via a Förster-type resonance energy transfer process from the BP in the excited triplet state to Rhodamine B acting as an acceptor fluorophore. This emission behavior was used for ratiometric afterglow sensing of water content in THF with a detection limit of 0.28 %, indicating that this study could pave the way for a new strategy for developing color-variable afterglow chemosensors for various analytes.
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BACKGROUND: Several effective therapies have been developed for spinal muscular atrophy (SMA), but there are multiple diseases that show SMA-like symptoms, necessitating efficient differential genetic diagnostic methods. Advancements in next-generation sequencing (NGS) technology have facilitated the successful diagnosis of many undiagnosed genetic diseases. Here, we applied NGS along with conventional methods for the molecular diagnosis of undiagnosed patients with lower motor neuron (LMN) symptoms who were initially suspected to have SMA. METHODS: We enrolled 157 patients with LMN symptoms who visited the Institute of Medical Genetics, Tokyo Women's Medical University, between 2005 and 2016. We excluded 86 patients diagnosed with SMA after confirming the causative SMN1 gene deletion or variants. Finally, we examined 12 undiagnosed patients from eight families by targeted resequencing using NGS. Variants were selected on the basis of literature search and databases, and mutations in a gene where loss of function is a known mechanism of disease were considered as pathogenic. Candidate variants were validated by Sanger sequencing. RESULTS: We detected novel variants for three patients from two families. Patients 1 and 2 (siblings) showed compound heterozygous TTN variants (c.6621delG, p.W2207Cfs*28 and c.23718T>A, p.F7906L), while patient 3 displayed compound heterozygous KIF1A variants of (c.3871C>T, p.R1291C and c.3898G>A, p.V1300M). CONCLUSIONS: We detected appropriate variants using our approach of obtaining candidate pathogenic variants by targeted resequencing through NGS and narrowed down the variants in light of patient clinical symptoms. We successfully identified novel causative variants in three undiagnosed patients, which indicated the effectiveness of our approach.
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Atrofia Muscular Espinal/genética , Atrofias Musculares Espinales de la Infancia/diagnóstico , Atrofias Musculares Espinales de la Infancia/genética , Niño , Conectina/genética , Conectina/metabolismo , Femenino , Eliminación de Gen , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Humanos , Cinesinas/genética , Cinesinas/metabolismo , Masculino , Atrofia Muscular Espinal/diagnóstico , Mutación , Proyectos Piloto , Eliminación de Secuencia , Proteína 1 para la Supervivencia de la Neurona Motora/genéticaRESUMEN
The dye-sensitized photoelectrochemical cells (DSPECs) incorporating a family of ruthenium complexes [RuII(bipyP)(bipy)2] (P1), [RuII(bipyP)(dmb)2] (P2), [RuII(bipyP)(dtbb)2] (P3) and [RuII(bipyP)(dnb)2] (P4), where bipyP = 2,2'-bipyridine-4,4'-diphosphonic acid, bipy = 2,2'-bipyridine, dmb = 4,4'-dimethyl-2,2'-bipyridine, dtbb = 4,4'-di-tert-butyl-2,2'-bipyridine, and dnb = 4,4'-dinonyl-2,2'-bipyridine, were fabricated in a dye-only system (do-DSPEC) and in a system where the electrolyte solution was loaded with EDTA sacrificial agent (sa-DSPEC). The increasing number of the alkyl chains of the ancillary bipy ligand shifts the ground- and excited-state potentials to the more negative values, although the introduction of the longer nonyl chain in P4 shows the opposite effect. In do-DSPECs, the photocurrent and hydrogen production performance follows the order P4 > P3 > P2 > P1, which correlates well with the degree of the excited-state quenching by electron injection to the conduction band of TiO2. The photoelectrochemistry of the sa-DSPECs reveals 10 times as many photocurrents as that measured in do-DSPECs, suggesting the ability of the hole to oxidize EDTA molecule. The hydrogen production performance of sa-DSPECs over five hours follows the order P2 > P1 > P3 > P4, which is consistent with the RuIII/RuII reorganization energies and the hole mobility on the TiO2 surface. The present study provides evidence that the subtle alkyl chain variation of the ruthenium photosensitizers can fine tune the electron injection capacity, RuIII/RuII self-exchange energetics and photostability of the complexes, which significantly influence the performance of the DSPECs.
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The self-assembly of polyvinyl alcohol (PVA) and benzene-1,4-diboronic acid (DBA) is employed as a sol-gel method for one-step fabrication of hydrogel coatings with versatile functionalities. A mixture of PVA and DBA in aqueous ethanol is prepared as a coating agent. The long pot life of the mixture allows for the coating of a wide range of materials with hydrogel films by simple cast- and dip-coating techniques. The resultant films show negligible dissolution in water and the intrinsic hydrophilicity of PVA provides the films with functional properties, such as improved antifogging property and resistance to protein and cell fouling. The self-assembling process shows adaptive inclusion properties toward nanoscale materials, such as metal-organic coordination polymers and inorganic nanoparticles, affording composite films. Furthermore, the coating film exhibits a unique secondary functionalization reactivity toward boronic acid-appended fluorescent dyes, through which a variety of materials are converted into fluorescent materials.
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Acetiltransferasas/genética , Bacillus subtilis/metabolismo , Proteínas Bacterianas/genética , Espermina/metabolismo , Acetiltransferasas/metabolismo , Bacillus subtilis/enzimología , Bacillus subtilis/genética , Proteínas Bacterianas/metabolismo , Medios de Cultivo/química , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Poliaminas/química , Poliaminas/metabolismo , Poliaminas/farmacología , Alimentos de Soja/microbiologíaRESUMEN
Chiral ensembles were spontaneously formed in solution through boronate esterification of structurally defined di(boronic acid)-appended tetraphenylethylene (DB-TPE) and commercially available l- or d-tartaric acid, showing enantioselective aggregation behavior for chiral diamines as well as cinchona alkaloids enabling the fluorescent recognition of their chirality.
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BACKGROUND: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder. Over 95% of SMA patients have homozygous deletions of the SMA-causative gene, SMN1. Thus, SMA carriers are usually diagnosed based on SMN1 copy number, with one copy indicating SMA carrier status. However, two SMN1 copies do not always exclude carrier status. In this study, we identified SMA carriers with two SMN1 copies. SUBJECTS AND METHODS: From 33 families, 65 parents of genetically confirmed SMA patients were tested to determine SMA carrier status. Molecular genetic analyses, including multiplex ligation-dependent probe amplification (MLPA) assay, were performed using blood samples from family members. RESULTS: Of the 65 parents, three parents from three families had two SMN1 copies. Accordingly, the frequency of carriers with two SMN1 copies was 4.6%. Two of these families were further studied. Patient 1 was homozygous for SMN1 deletion. Patient 1's mother had two SMN1 copies on one chromosome, with deletion of SMN1 on the other chromosome ([2+0] genotype). Patient 1 inherited SMN1-deleted chromosomes from both parents. Patient 2 was compound heterozygous for two SMN1 mutations: whole-gene deletion and intragenic missense mutation, c.826T>C (p.Tyr276His). Patient 2's father had two SMN1 copies with the same intragenic mutation in one copy ([1+1d] genotype, d intragenic mutation). Patient 2 inherited the chromosome with an SMN1 mutation from the father and SMN1-deleted chromosome from the mother. CONCLUSION: SMA carriers with two SMN1 copies may be rare, but its possibility should be taken into consideration in carrier testing and counseling for SMA families or population-based carrier screening.
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Atrofia Muscular Espinal/genética , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Adulto , Variaciones en el Número de Copia de ADN , Familia , Femenino , Eliminación de Gen , Tamización de Portadores Genéticos/métodos , Asesoramiento Genético , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa Multiplex/métodos , Atrofia Muscular Espinal/metabolismo , Mutación/genética , Linaje , Eliminación de Secuencia/genética , Proteína 1 para la Supervivencia de la Neurona Motora/sangre , Proteína 1 para la Supervivencia de la Neurona Motora/metabolismoRESUMEN
Dye-sensitized photoelectrochemical cells (DSPECs) composed of a new near-infrared BODIPY dye D1 that is co-deposited with a ruthenium water oxidation catalyst C1 have been fabricated. The devices at pH 7.2 showed an excellent Faradaic efficiency of H2 production (65.8%) that was 5.4 times larger than that of a triphenylamine photosensitizer D2 and C1-coadsorbed cell.
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Surface modification of a polyvinyl alcohol sponge with functionalized boronic acids led to the formation of porous materials applicable for multicolor emission, chemical sensing and 3D cell culture.
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Polyamines have beneficial properties to prevent aging-associated diseases. Raw soybean has relatively high polyamine contents; and the fermented soybean natto is a good source of polyamines. However, detailed information of diversity of polyamine content in raw soybean is lacking. The objectives of this study were to evaluate differences of polyamines among raw soybeans and select the high polyamine-containing cultivar for natto production. Polyamine contents were measured chromatographically in 16 samples of soybean, which showed high variation among soybeans as follows: 93-861 nmol/g putrescine, 1055-2306 nmol/g spermidine, and 177-578 nmol/g spermine. We then confirmed the high correlations of polyamine contents between raw soybean and natto (r = 0.96, 0.95, and 0.94 for putrescine, spermidine, and spermine, respectively). Furthermore, comparison of the polyamine contents among 9 Japanese cultivars showed that 'Nakasen-nari' has the highest polyamine contents, suggesting its suitability for enhancement of polyamine contents of natto.
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Glycine max/química , Glycine max/genética , Poliaminas/análisis , Alimentos de Soja/análisis , Bacillus subtilis/fisiología , Análisis de los Alimentos , Japón , Alimentos de Soja/microbiologíaRESUMEN
Self-assembled colorimetric sensors have been prepared from Zn(II)-DPA-attached phenylboronic acid (·Zn) and catechol-type dyes. The ·Zn-dye sensors display selectivity towards oligophosphate over monophosphates. The colorimetric sensor assay (·Zn-dye) is utilized to monitor a model of a metabolic reaction where ATP is hydrolyzed to pyrophosphate (PPi) and AMP.
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Adenosina Trifosfato/análisis , Adenosina Trifosfato/metabolismo , Colorimetría , Modelos Biológicos , Agua/química , Adenosina Monofosfato/química , Adenosina Monofosfato/metabolismo , Difosfatos/química , Difosfatos/metabolismo , HidrólisisRESUMEN
BACKGROUND: Most patients with spinal muscular atrophy lack the survival motor neuron 1 gene (SMN1) in the telomeric region of the spinal muscular atrophy locus on chromosome 5q13. On the other hand, the copy number of SMN2, a centromeric homolog of SMN1, is increased in many of these patients. This study aimed to clarify the mechanism underlying these structural variations. METHODS: We determined the copy numbers of telomeric and centromeric genes in the spinal muscular atrophy locus of 86 patients and 22 control subjects using multiplex ligation-dependent probe amplification analysis. Then, we chose 74 patients lacking SMN1 exons 7 and 8, and compared their dataset with that of 22 control subjects retaining SMN1 exons 7 and 8. RESULTS: The SMN2 copy number was shown to vary widely and to correlate with the disease severity of the patients. Interestingly, telomeric NAIP and telomeric GTF2H2 showed similar tendencies. We also noted positive correlations among the copy number of SMN2 and the telomeric genes of the spinal muscular atrophy locus. However, the copy numbers of centromeric NAIP and centromeric GTF2H2 were stable among the patients, with both approximating a value of two. CONCLUSION: Our findings suggested that the telomeric region of the spinal muscular atrophy locus appears to be susceptible to structural variation, whereas the centromeric region is stable. Moreover, according to our results, new SMN2 copies may be generated in the telomeric region of the spinal muscular atrophy locus, supporting the SMN1-to-SMN2 gene conversion theory.