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Octacalcium phosphate (OCP), an inorganic compound with a layered structure that can incorporate various carboxylic acids, exhibits fluorescence when aromatic carboxylate ions are present in the interlayers. However, the incorporation of carboxylate ions into OCP involves molecular selectivity, and the synthesis of novel OCP materials with incorporated terephthalate ion derivatives is particularly challenging. In this study, we incorporated 4-(carboxymethyl)benzoate and 1,4-phenylenediacetate ions into OCP for the first time and investigated the resulting fluorescence properties. The relationship between the (100) interplanar spacing and size of the incorporated dicarboxylate ions revealed that 4-(carboxymethyl)benzoate ions have a relaxed structure in the OCP interlayers, whereas the structure of the 1,4-phenylenediacetate ions is elongated by approximately 10% relative to that of the stable conformation. OCP with incorporated 1,4-phenylenediacetate ions showed blue fluorescence at 286 nm under 254 nm excitation. In contrast, distinct from previously reported fluorescent OCPs, OCP with incorporated 4-(carboxymethyl)benzoate ions exhibited two-colour fluorescence, with pink emission under 254 nm excitation and blue emission under 312 and 365 nm excitation. This OCP material exhibiting fluorescence at two wavelengths in the visible-light range offers new possibilities for practical applications. In particular, these unique fluorescence characteristics combined with the excellent biological properties of OCP can be exploited to develop novel biofriendly fluorescent probes. These findings contribute to an improved understanding of fundamental calcium phosphate chemistry and should encourage further research on functional OCP materials.
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Melanin is produced by melanocytes to protect human skin from harmful ultraviolet radiation. During skin cell renewal, melanin and dead skin cells are disposed of. However, prolonged exposure to ultraviolet rays or aging can disturb this cycle, leading to skin hyperpigmentation due to melanin accumulation. Tyrosinase is a crucial enzyme involved in melanin biosynthesis. Although various compounds, including tyrosine inhibitors, that counteract melanin accumulation have been reported, some, such as hydroquinone, are toxic and can cause vitiligo. Meanwhile, the skin is the largest organ and the outermost layer of the immune system, containing a diverse range of bacteria that produce low-toxicity compounds. In the current study, we aim to identify metabolites produced by skin microbiota that inhibit tyrosinase. Specifically, mushroom tyrosinase served as the study model. Following commensal skin bacteria screening, Corynebacterium tuberculostearicum was found to inhibit tyrosinase activity. The active compound was cyclo(l-Pro-l-Tyr); commercially available cyclo(l-Pro-l-Tyr) also exhibited inhibitory activity. Docking simulations suggested that cyclo(l-Pro-l-Tyr) binds to the substrate-binding site of mushroom tyrosinase, obstructing the substrate pocket and preventing its activity. Hence, cyclo(l-Pro-l-Tyr) might have potential applications as a cosmetic agent and food additive.
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Corynebacterium , Monofenol Mono-Oxigenase , Pele , Monofenol Mono-Oxigenase/antagonistas & inibidores , Monofenol Mono-Oxigenase/metabolismo , Humanos , Pele/microbiologia , Pele/efeitos dos fármacos , Pele/metabolismo , Simulação de Acoplamento Molecular , Agaricales/enzimologia , Inibidores Enzimáticos/farmacologia , Peptídeos Cíclicos/farmacologia , Peptídeos Cíclicos/química , Melaninas/metabolismo , Melaninas/biossínteseRESUMO
Xyloglucan in plant cell walls has complex side-chain structures; Aspergillus oryzae produces various enzymes to degrade and assimilate xyloglucan. In this study, we identified and characterized α-1,2-l-fucosidase (AfcA) which is involved in xyloglucan degradation in A. oryzae. AfcA expression was induced in the presence of xyloglucan oligosaccharides. AfcA showed specific activity toward α-(1â2)-linked l-fucopyranosyl residues attached to the side chains of xyloglucan oligosaccharides and milk oligosaccharides, but not toward α-(1â3)-, α-(1â4)-, and α-(1â6)-linked l-fucopyranosyl residues. As fucopyranosyl residues in the side chains of xyloglucan oligosaccharides prevent the degradation of xyloglucan oligosaccharides by isoprimeverose-producing oligoxyloglucan hydrolase and ß-galactosidase, the cooperative action of AfcA, isoprimeverose-producing oligoxyloglucan hydrolase, and ß-galactosidase play a key role in degrading fucosylated xyloglucan in A. oryzae.
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Aspergillus oryzae , Glucanos , Xilanos , alfa-L-Fucosidase , Xilanos/metabolismo , Xilanos/química , Glucanos/metabolismo , Glucanos/química , Aspergillus oryzae/enzimologia , Aspergillus oryzae/metabolismo , alfa-L-Fucosidase/metabolismo , alfa-L-Fucosidase/química , alfa-L-Fucosidase/genética , Oligossacarídeos/metabolismo , Oligossacarídeos/química , beta-Galactosidase/metabolismo , beta-Galactosidase/química , Especificidade por Substrato , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Parede Celular/metabolismo , DissacarídeosRESUMO
Changes in the sea surface temperature (SST) pattern in the tropical Pacific modulate radiative feedbacks to greenhouse gas forcing, the pace of global warming and regional climate impacts. Therefore, elucidating the drivers of the pattern is critically important for reducing uncertainties in future projections. However, the causes of observed changes over recent decades, an enhancement of the zonal SST contrast coupled with a strengthening of the Walker circulation, are still debated. Here we focus on the role of external forcing and review existing mechanisms of the forced response categorized as either an energy perspective that adopts global and hemispheric energy budget constraints or a dynamical perspective that examines the atmosphere-ocean coupled processes. We then discuss their collective and relative contributions to the past and future SST pattern changes and propose a narrative that reconciles them. Although definitive evidence is not yet available, our assessment suggests that the zonal SST contrast has been dominated by strengthening mechanisms in the past, but will shift towards being dominated by weakening mechanisms in the future. Finally, we present opportunities to resolve the model-observations discrepancy regarding the recent trends.
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Acetogens are among the key microorganisms involved in the bioproduction of commodity chemicals from diverse carbon resources, such as biomass and waste gas. Thermophilic acetogens are particularly attractive because fermentation at higher temperatures offers multiple advantages. However, the main target product is acetic acid. Therefore, it is necessary to reshape metabolism using genetic engineering to produce the desired chemicals with varied carbon lengths. Although such metabolic engineering has been hampered by the difficulty involved in genetic modification, a model thermophilic acetogen, M. thermoacetica ATCC 39073, is the case with a few successful cases of C2 and C3 compound production, other than acetate. This brief report attempts to expand the product spectrum to include C4 compounds by using strain Y72 of Moorella thermoacetica. Strain Y72 is a strain related to the type strain ATCC 39073 and has been reported to have a less stringent restriction-modification system, which could alleviate the cumbersome transformation process. A simplified procedure successfully introduced a key enzyme for acetoin (a C4 chemical) production, and the resulting strains produced acetoin from sugars and gaseous substrates. The culture profile revealed varied acetoin yields depending on the type of substrate and culture conditions, implying the need for further engineering in the future. Thus, the use of a user-friendly chassis could benefit the genetic engineering of M. thermoacetica.
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Genome Editing is widely used in biomedical research and medicine. Zinc finger nucleases (ZFNs) are smaller in size than transcription activator-like effector (TALE) nucleases (TALENs) and CRISPR-Cas9. Therefore, ZFN-encoding DNAs can be easily packaged into a viral vector with limited cargo space, such as adeno-associated virus (AAV) vectors, for in vivo and clinical applications. ZFNs have great potential for translational research and clinical use. However, constructing functional ZFNs and improving their genome editing efficiency is extremely difficult. Here, the efficient construction of functional ZFNs and the improvement of their genome editing efficiency using AlphaFold, Coot, and Rosetta are described. Plasmids encoding ZFNs consisting of six fingers using publicly available zinc-finger resources are assembled. Two functional ZFNs from the ten ZFNs tested are successfully obtained. Furthermore, the engineering of ZFNs using AlphaFold, Coot, or Rosetta increases the efficiency of genome editing by 5%, demonstrating the effectiveness of engineering ZFNs based on structural modeling.
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Edição de Genes , Nucleases de Dedos de Zinco , Edição de Genes/métodos , Nucleases de Dedos de Zinco/genética , Nucleases de Dedos de Zinco/metabolismo , Humanos , Dedos de Zinco/genética , Sistemas CRISPR-Cas/genéticaRESUMO
[Purpose] The sense of vision is omitted in blind soccer, and sound source localization to grasp the position of the ball is extremely important. The purpose of this study was to clarify whether there is a difference in ability in sound source localization in its approaching condition between visually impaired and sighted people, using the source actually used in blind soccer ball competitions. [Participants and Methods] Eighteen participants were divided into two groups; 10 sighted people and eight visually impaired people. The participants were asked to press a switch when a rolling blind soccer ball was sensed in any one of the four directions. We recorded time error as the difference between the time when the ball passed the optical sensor set under the participant's feet and when the participant pressed the switch. [Results] The time error in response increased with the ball speed in all cases; however, its dependence on the ball speed was significantly different between the two groups. [Conclusion] The visually impaired participants made less time errors in response to the localization of the ball than the sighted participants, even when the ball speed increased. The results indicate that visually impaired people have better sound source localization ability than sighted people do.
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Antimicrobial-product package inserts and insufficient staffing impede routine carbapenem monitoring in the inpatient setting in Japan. The collaboration between antimicrobial stewardship teams and clinical pharmacists was associated with a sustained improvement in carbapenem dosing optimization. Our findings could be of use to countries with inadequate monitoring of carbapenem antimicrobial use.
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Porphyromonas gingivalis, a Gram-negative anaerobic bacillus, is the primary pathogen in periodontitis. Herein, we cultivated strains of oral bacteria, including P. gingivalis and the oral commensal bacteria Actinomyces viscosus and Streptococcus mutans, and recorded the infrared absorption spectra of the gases released by the cultured bacteria at a resolution of 0.5 cm-1 within the wavenumber range of 500-7500 cm-1. From these spectra, we identified the infrared wavenumbers associated with characteristic absorptions in the gases released by P. gingivalis using a decision tree-based machine learning algorithm. Finally, we compared the obtained absorbance spectra of ammonia (NH3) and carbon monoxide (CO) using the HITRAN database. We observed peaks at similar positions in the P. gingivalis gases, NH3, and CO spectra. Our results suggest that P. gingivalis releases higher amounts of NH3 and CO than A. viscosus and S. mutans. Thus, combining Fourier transform infrared spectroscopy with machine learning enabled us to extract the specific wavenumber range that differentiates P. gingivalis from a vast dataset of peak intensity ratios. Our method distinguishes the gases from P. gingivalis from those of other oral bacteria and provides an effective strategy for identifying P. gingivalis in oral bacteria. Our proposed methodology could be valuable in clinical settings as a simple, noninvasive pathogen diagnosis technique.
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Periodontite , Porphyromonas gingivalis , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Periodontite/microbiologia , GasesRESUMO
BACKGROUND: Isopropanol (IPA) is a commodity chemical used as a solvent or raw material for polymeric products, such as plastics. Currently, IPA production depends largely on high-CO2-emission petrochemical methods that are not sustainable. Therefore, alternative low-CO2 emission methods are required. IPA bioproduction using biomass or waste gas is a promising method. RESULTS: Moorella thermoacetica, a thermophilic acetogenic microorganism, was genetically engineered to produce IPA. A metabolic pathway related to acetone reduction was selected, and acetone conversion to IPA was achieved via the heterologous expression of secondary alcohol dehydrogenase (sadh) in the thermophilic bacterium. sadh-expressing strains were combined with acetone-producing strains, to obtain an IPA-producing strain. The strain produced IPA as a major product using hexose and pentose sugars as substrates (81% mol-IPA/mol-sugar). Furthermore, IPA was produced from CO, whereas acetate was an abundant byproduct. Fermentation using syngas containing both CO and H2 resulted in higher IPA production at the specific rate of 0.03 h-1. The supply of reducing power for acetone conversion from the gaseous substrates was examined by supplementing acetone to the culture, and the continuous and rapid conversion of acetone to IPA showed a sufficient supply of NADPH for Sadh. CONCLUSIONS: The successful engineering of M. thermoacetica resulted in high IPA production from sugars. M. thermoacetica metabolism showed a high capacity for acetone conversion to IPA in the gaseous substrates, indicating acetone production as the bottleneck in IPA production for further improving the strain. This study provides a platform for IPA production via the metabolic engineering of thermophilic acetogens.
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A fetal growth restriction is related to adverse child outcomes. We investigated risk ratios and population-attributable fractions (PAF) of small-for-gestational-age (SGA) infants in the Japanese population. Among 28,838 infants from five ongoing prospective birth cohort studies under the Japan Birth Cohort Consortium, two-stage individual-participant data meta-analyses were conducted to calculate risk ratios and PAFs for SGA in advanced maternal age, pre-pregnancy underweight, and smoking and alcohol consumption during pregnancy. Risk ratio was calculated using modified Poisson analyses with robust variance and PAF was calculated in each cohort, following common analyses protocols. Then, results from each cohort study were combined by meta-analyses using random-effects models to obtain the overall estimate for the Japanese population. In this meta-analysis, an increased risk (risk ratio, [95% confidence interval of SGA]) was significantly associated with pre-pregnancy underweight (1.72 [1.42-2.09]), gestational weight gain (1.95 [1.61-2.38]), and continued smoking during pregnancy (1.59 [1.01-2.50]). PAF of underweight, inadequate gestational weight gain, and continued smoking during pregnancy was 10.0% [4.6-15.1%], 31.4% [22.1-39.6%], and 3.2% [-4.8-10.5%], respectively. In conclusion, maternal weight status was a major contributor to SGA births in Japan. Improving maternal weight status should be prioritized to prevent fetal growth restriction.
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Retardo do Crescimento Fetal , Ganho de Peso na Gestação , Criança , Lactente , Feminino , Gravidez , Humanos , Retardo do Crescimento Fetal/epidemiologia , Japão/epidemiologia , Coorte de Nascimento , Estudos de Coortes , Estudos Prospectivos , MagrezaRESUMO
We previously found that ribosomal protein L9 (RPL9) is a novel advanced glycation end product (AGE)-binding protein that can decrease pro-inflammatory TNF-α expression stimulated by lipopolysaccharide (LPS) plus high-mobility group box 1 (HMGB1), suggesting that RPL9 has a role in regulating LPS+HMGB1-stimulated inflammatory reactions. Among the various ribosomal proteins, it was found that RPS5 reproduced the regulatory activity of RPL9 on LPS+HMGB1-stimulated TNF-α expression in macrophage-like RAW264.7 cells. RPL9 and RPS5 share a common feature as cationic proteins. Polylysine, a cationic polypeptide, and a synthetic peptide of the cationic region from RPL9 also exhibited reducing activity on LPS+HMGB1-induced TNF-α expression. By pull-down assay, RPL9 and RPS5 were confirmed to interact with AGEs. When AGEs coexisted with LPS, HMGB1, plus RPL9 or RPS5, the reducing effect of TNF-α expression by these cationic ribosomal proteins was shown to be abrogated. The results suggest that cationic ribosomal proteins have a regulatory role in the pro-inflammatory response induced by LPS+HMGB1, and in the pathophysiological condition of accumulating AGEs, this regulatory effect is abolished, which exacerbates inflammation.
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Proteína HMGB1 , Lipopolissacarídeos , Humanos , Lipopolissacarídeos/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Ribossômicas , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Produtos Finais de Glicação AvançadaRESUMO
Fatty acid-binding protein 7 (FABP7) is vital for uptake and trafficking of fatty acids in the nervous system. To investigate the involvement of FABP7 in noise-induced hearing loss (NIHL) pathogenesis, we used Fabp7 knockout (KO) mice generated via CRISPR/Cas9 in the C57BL/6 background. Initial auditory brainstem response (ABR) measurements were conducted at 9 weeks, followed by noise exposure at 10 weeks. Subsequent ABRs were performed 24 h later, with final measurements at 12 weeks. Inner ears were harvested 24 h after noise exposure for RNA sequencing and metabolic analyses. We found no significant differences in initial ABR measurements, but Fabp7 KO mice showed significantly lower thresholds in the final ABR measurements. Hair cell survival was also enhanced in Fabp7 KO mice. RNA sequencing revealed that genes associated with the electron transport chain were upregulated or less impaired in Fabp7 KO mice. Metabolomic analysis revealed various alterations, including decreased glutamate and aspartate in Fabp7 KO mice. In conclusion, FABP7 deficiency mitigates cochlear damage following noise exposure. This protective effect was supported by the changes in gene expression of the electron transport chain, and in several metabolites, including excitotoxic neurotransmitters. Our study highlights the potential therapeutic significance of targeting FABP7 in NIHL.
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Perda Auditiva Provocada por Ruído , Audição , Camundongos , Animais , Proteína 7 de Ligação a Ácidos Graxos/genética , Proteína 7 de Ligação a Ácidos Graxos/metabolismo , Camundongos Endogâmicos C57BL , Audição/fisiologia , Ruído/efeitos adversos , Perda Auditiva Provocada por Ruído/genética , Cóclea/metabolismo , Camundongos Knockout , Potenciais Evocados Auditivos do Tronco Encefálico/fisiologia , Limiar Auditivo/fisiologiaRESUMO
Mannosylerythritol lipids (MELs) are extracellular glycolipids produced by the basidiomycetous yeast strains. MELs consist of the disaccharide mannosylerythritol, which is acylated with fatty acids and acetylated at the mannose moiety. In the MEL biosynthesis pathway, an acyltransferase from Pseudozyma tsukubaensis, PtMAC2p, a known excellent MEL producer, has been identified to catalyze the acyl-transfer of fatty acid to the C3'-hydroxyl group of mono-acylated MEL; however, its structure remains unclear. Here, we performed X-ray crystallography of recombinant PtMAC2p produced in Escherichia coli and homogeneously purified it with catalytic activity in vitro. The crystal structure of PtMAC2p was determined by single-wavelength anomalous dispersion using iodide ions. The crystal structure shows that PtMAC2p possesses a large putative catalytic tunnel at the center of the molecule. The structural comparison demonstrated that PtMAC2p is homologous to BAHD acyltransferases, although its amino acid-sequence identity was low (<15%). Interestingly, the HXXXD motif, which is a conserved catalytic motif in the BAHD acyltransferase superfamily, is partially conserved as His158-Thr159-Leu160-Asn161-Gly162 in PtMAC2p, i.e., D in the HXXXD motif is replaced by G in PtMAC2p. Site-directed mutagenesis of His158 to Ala resulted in more than 1,000-fold decrease in the catalytic activity of PtMAC2p. These findings suggested that His158 in PtMAC2p is the catalytic residue. Moreover, in the putative catalytic tunnel, hydrophobic amino acid residues are concentrated near His158, suggesting that this region is a binding site for the fatty acid side chain of MEL (acyl acceptor) and/or acyl-coenzyme A (acyl donor). To our knowledge, this is the first study to provide structural insight into the catalytic activity of an enzyme involved in MEL biosynthesis.
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Advanced glycation end products (AGEs) are a heterogeneous group of compounds that are non-enzymatically produced by reactions between carbonyl compounds and proteins. Many types of AGEs are produced according to the type or concentration of the reacting carbonyl compound. We have previously demonstrated that a glycolaldehyde-derived AGE suppresses stimulator of interferon gene (STING)/TANK-binding kinase 1 (TBK1)/interferon regulatory transcription factor 3 (IRF3), which is a component of the innate immune system. In this report, we investigated the effects of AGEs prepared by several carbonyl compounds on STING/TBK1/IRF3 signaling. AGEs used in the present study were numbered based on the carbonyl compound type: AGE1, derived from glucose; AGE2, derived from glyceraldehyde; AGE3, derived from glycolaldehyde; AGE4, derived from methylglyoxal; and AGE5, derived from glyoxal. AGEs derived from aldehyde (AGE2 and AGE3) and dicarbonyl compounds (AGE4 and AGE5) suppressed cyclic GMP-AMP (cGAMP)-induced activation of STING/TBK1/IRF3 signaling, with different suppression efficiencies observed. Lysine modification by carbonyl compounds was related to the efficiency of the suppressive effect on STING/TBK1/IRF3 signaling. Among the AGEs used, only AGE1 enhanced cGAMP-induced activation of STING/TBK1/IRF3 signaling. Enhancing the modulation of STING/TBK1/IRF3 signaling by AGE1 was mediated by toll-like receptor 4. These results indicated that modulation of STING/TBK1/IRF3 signaling by prepared AGEs is dependent on the type and concentration of the carbonyl compound present. Modulating STING/TBK1/IRF3 signaling by AGEs may involve modification of lysine residues in proteins.
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Lisina , Proteínas de Membrana , Fosforilação , Lisina/metabolismo , Proteínas de Membrana/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Interferons/metabolismoRESUMO
Niemann-Pick disease type C (NPC) is an autosomal recessive disorder with progressive neurodegeneration. Although the causative genes were previously identified, NPC has unclear pathophysiological aspects, and patients with NPC present various symptoms and onset ages. However, various novel biomarkers and metabolic alterations have been investigated; at present, few comprehensive proteomic alterations have been reported in relation to NPC. In this study, we aimed to elucidate proteomic alterations in NPC and perform a global proteomics analysis for NPC model cells. First, we developed two NPC cell models by knocking out NPC1 using CRISPR/Cas9 (KO1 and KO2). Second, we performed a label-free (LF) global proteomics analysis. Using the LF approach, more than 300 proteins, defined as differentially expressed proteins (DEPs), changed in the KO1 and/or KO2 cells, while the two models shared 35 DEPs. As a bioinformatics analysis, the construction of a protein-protein interaction (PPI) network and an enrichment analysis showed that common characteristic pathways such as ferroptosis and mitophagy were identified in the two model cells. There are few reports of the involvement of NPC in ferroptosis, and this study presents ferroptosis as an altered pathway in NPC. On the other hand, many other pathways and DEPs were previously suggested to be associated with NPC, supporting the link between the proteome analyzed here and NPC. Therapeutic research based on these results is expected in the future.
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Doença de Niemann-Pick Tipo C , Humanos , Doença de Niemann-Pick Tipo C/metabolismo , Proteômica/métodos , Proteoma , Hepatócitos/metabolismoRESUMO
Recently, prediction of gestational diabetes mellitus (GDM) using artificial intelligence (AI) from medical records has been reported. We aimed to evaluate GDM-predictive AI-based models using birth cohort data with a wide range of information and to explore factors contributing to GDM development. This investigation was conducted as a part of the Japan Environment and Children's Study. In total, 82,698 pregnant mothers who provided data on lifestyle, anthropometry, and socioeconomic status before pregnancy and the first trimester were included in the study. We employed machine learning methods as AI algorithms, such as random forest (RF), gradient boosting decision tree (GBDT), and support vector machine (SVM), along with logistic regression (LR) as a reference. GBDT displayed the highest accuracy, followed by LR, RF, and SVM. Exploratory analysis of the JECS data revealed that health-related quality of life in early pregnancy and maternal birthweight, which were rarely reported to be associated with GDM, were found along with variables that were reported to be associated with GDM. The results of decision tree-based algorithms, such as GBDT, have shown high accuracy, interpretability, and superiority for predicting GDM using birth cohort data.
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Diabetes Gestacional , Gravidez , Feminino , Humanos , Criança , Diabetes Gestacional/diagnóstico , Diabetes Gestacional/epidemiologia , Coorte de Nascimento , Inteligência Artificial , Japão/epidemiologia , Qualidade de Vida , Mães , Aprendizado de MáquinaRESUMO
Octacalcium phosphate (OCP) is a layered compound capable of incorporating carboxylate ions within its interlayer structure. In this study, we successfully synthesised OCP with incorporated 3,3'-dithiodipropionate ions. Our finding is beneficial for the development of novel OCP-based materials with dynamic properties derived from disulfide bonds.