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Antibiotic-associated diarrhea is a relatively common problem, and the main bacterial cause is Clostridioides difficile followed by Staphylococcus aureus and other pathogens. The diagnostic procedure for methicillin-resistant S. aureus enteritis is not well established. Phagocytosis is a key antimicrobial process involved in host defense. Phagocytosed bacteria identified by Gram staining are one marker to identify causative microorganisms and select subsequent treatment strategies. However, there are few reports on fecal Gram staining using phagocytosed bacteria as a target for diarrhea treatment. We report the successful use of fecal Gram staining to diagnose and treat methicillin-resistant S. aureus enteritis.

We present the spectroscopic discovery of 69 quasars at 5.8 < z < 7.0, drawn from the Hyper Suprime-Cam (HSC) Subaru Strategic Program (SSP) imaging survey data. This is the 16th publication from the Subaru High-z Exploration of Low-Luminosity Quasars (SHELLQs) project, and completes identification of all but the faintest candidates (i.e., i-band dropouts with zAB < 24 and y-band detections, and z-band dropouts with yAB < 24) with Bayesian quasar probability Pq > 0.1 in the HSC-SSP third public data release (PDR3). The sample reported here also includes three quasars with Pq < 0.1 at z ~ 6.6, which we selected in an effort to completely cover the reddest point sources with simple color cuts. The number of high-z quasars discovered in SHELLQs has now grown to 162, including 23 type-II quasar candidates. This paper also presents identification of seven galaxies at 5.6 < z < 6.7, an [O III] emitter at z = 0.954, and 31 Galactic cool stars and brown dwarfs. High-z quasars and galaxies comprise 75 % and 16 % respectively of all the spectroscopic SHELLQs objects that pass our latest selection algorithm with the PDR3 photometry. That is, a total of 91 % of the objects lie at z > 5.6. This demonstrates that the algorithm has very high efficiency, even though we are probing an unprecedentedly low-luminosity population down to M1450 ~ -21 mag., Accepted for publication in The Astrophysical Journal Supplement Series

Devido à Pandemia Covid-19, no Brasil, as atividades presenciais dos cursos de medicina e ligas acadêmicas foram suspensas, ambas enfrentaram dificuldades para readaptar suas atividades ao formato remoto/virtual. O presente artigo objetiva relatar a experiência de atuação da Liga de Anatomia Clínica e Cirúrgica da Universidade Federal do Paraná, Campus Toledo, com ênfase na adaptação das atividades presenciais ao ambiente remoto/virtual de ensino e nas contribuições ao aprendizado da anatomia, na pandemia. Trata-se de um estudo descritivo, transversal, retrospectivo de caráter qualitativo, um relato de experiência. Realizou-se, na pandemia, inúmeras atividades em plataformas online, aulas quinzenais, majoritariamente abertas ao público e certificadas, postagens de materiais revisionais no Instagram da liga, como “AnatoCards” e divulgação de aulas/eventos. Realizou-se, de modo on-line, a Primeira Jornada de Anatomia Clínica e Cirúrgica do Oeste do Paraná, com palestrantes de diferentes especialidades médicas e regiões do país, e alcance superior a 2500 pessoas. Criação do “Anatodúvidas”, em 2021, no Instagram, projeto de extensão da liga, que democratizou o acesso ao conhecimento anatômico, embasado cientificamente, em linguagem acessível, à comunidade. Efetuou-se uma matéria em Jornal da Região para ampliar a visibilidade da liga para milhares de pessoas. O incentivo à pesquisa resultou em duas publicações de resumos em Congressos de Medicina. Portanto, a atuação da liga na pandemia contribuiu ao amparo dos pilares ensino, pesquisa e extensão, e estimulou o engajamento dos discentes. Isso resultou em ampla promoção, extensão e aprimoramento dos conhecimentos anatômicos, contribuindo na formação médica mais humana e completa, beneficiando amplamente à comunidade.

Nuclear factor (NF)-κB essential modifier (NEMO), also known as IκB kinase subunit-γ (IKKγ), is a pivotal molecule in the NF-κB signaling pathway. Mutations of NEMO cause incontinentia pigmenti and X-linked ectodermal dysplasia with immunodeficiency. Mendelian susceptibility to mycobacterial diseases (MSMD), which confers an almost selective predisposition to mycobacterial infection, is also caused by NEMO mutations. We herein report the first case of a patient with X-linked recessive (XR) MSMD who developed cutaneous squamous cell carcinoma, thyroid cancer and Langerhans cell histiocytosis. The relationship between NEMO mutation and oncogenesis is discussed.

The purpose of this study was to establish a method for determining the bacteriolytic activity after separation of lysozyme-binding proteins from egg white. Lysozyme-binding proteins such as ovotransferrin and ovalbumin were separated by non-denaturing two-dimensional electrophoresis (2DE) and transferred to a membrane. The lysozyme activity of the separated and immobilized egg white proteins was assessed directly to produce a non-denaturing 3D map of the egg white proteins by incorporating an axis that combined each spot's lysozyme-activity with the non-denaturing 2DE pattern. Lysozyme-ovotransferrin and lysozyme-ovalbumin complexes could be reconstructed in vitro after the cathode end fraction containing lysozyme was added to purified ovotransferrin and ovalbumin, respectively. These complexes retained lysozyme activity even after separation by non-denaturing 2DE. Furthermore, when the lysozyme-ovotransferrin complex from egg white was extracted after separation by isoelectric focusing by replacing the cathodic sodium hydroxide solution with phosphoric acid solution, the complex possessed bacteriolytic activity against both Bacillus subtilis and Escherichia coli. These methods can be applied to investigate protein complexes possessing bacteriolytic activity against a wide range of both Gram-positive and Gram-negative bacteria.

Genetically encoded calcium indicators (GECIs) are suitable for long-term imaging studies. In this study, we employed a highly sensitive GECI, G-GECO, and achieved efficient gene delivery with an adenoviral vector. The adenoviral vector allowed us to express G-GECO in more than 80% of cells. More than 80% of G-GECO-expressing cells showed an ATP-induced increase in fluorescence intensity due to Ca2+ release from intracellular stores and subsequent Ca2+ entry. The fluorescence intensity of these cells was increased more than 2-fold by stimulation with 10 μM ATP. We applied long-term imaging (for ~10 h) to monitor Ca2+ responses in SF2, a rat dental epithelial cell line, in culture conditions. SF2 cells showed intermittent rises in the intracellular Ca2+ concentration in the presence of 100 nM 1,25-dihydroxyvitamin D3. Many of these Ca2+ responses began at a specific location in the cytoplasm and spread throughout the entire cytoplasm. The combination of efficient gene delivery with an adenoviral vector and long-term imaging with a highly sensitive GECI enabled detection of intermittent Ca2+ responses that occur only 3-10 times/h/100 cells. This method could be useful to study the effects of Ca2+ responses for regulating longterm processes, such as gene expression, cell migration, and cell division, in many cell types.

Recent studies cite β2-adrenergic receptor (β2AR) antagonists as novel therapeutic agents for melanoma, as they may reduce the disease progression. The β2AR has shown to be expressed in malignant melanoma. However, it remains unclear whether the β2AR expression has a clinical and pathological significance in patients with cutaneous malignant melanoma. We herein conducted a clinicopathological study to investigate the protein expression of β2AR in malignant melanoma of the skin and its prognostic significance. One hundred thirty-three patients with surgically resected cutaneous malignant melanoma were evaluated. Tumor sections were stained by immunohistochemistry for β2AR, Ki-67, the microvessel density (MVD) determined by CD34, and p53. β2AR was highly expressed in 44.4 % (59 out of 133) of the patients. The expression of β2AR was significantly associated with the tumor thickness, ulceration, T factor, N factor, disease stage, tumor size, cell proliferation (Ki-67), and MVD (CD34). Using Spearman's rank test, the β2AR expression was correlated with Ki-67 (r = 0.278; 95 % CI, 0.108 to 0.432; P = 0.001), CD34 (r = 0.445; 95 %CI, 0.293 to 0.575; P

In this study, the combined use of a biopolymer chitosan and an oxidoreductase polyphenol oxidase (PPO) was systematically investigated for the removal of bisphenol derivatives from aqueous medium. The process parameters, such as the pH value, temperature, and PPO concentration, were estimated to conduct the enzymatic quinone oxidation of bisphenol derivatives by as little enzyme as possible. Bisphenol derivatives effectively underwent PPO-catalysed quinone oxidation without H2O2 unlike other oxidoreductases, such as peroxidase and tyrosinase, and the optimum conditions were determined to be pH 7.0 and 40°C for bisphenol B, bisphenol E, bisphenol O, and bisphenol Z; pH 7.0 and 30°C for bisphenol C and bisphenol F; and pH 8.0 and 40°C for bisphenol T. They were completely removed through adsorption of enzymatically generated quinone derivatives on chitosan beads or chitosan powders. Quinone adsorption on chitosan beads or chitosan powders in the heterogeneous system was found to be a more effective procedure than generation of aggregates in the homogeneous system with chitosan solution. The removal time was shortened by increasing the amount of chitosan beads or decreasing the size of the chitosan powders.

Study of prehistoric to medieval-age tsunami deposits along a riverbank site near the eastern Nankai Trough, central Japan, show that, not only did Tokai earthquakes occur with a higher frequency than previously thought, but that contemporaneous ruptures of the Tokai and Nankai fault segments were also more common. The site revealed a ∼1-km long coast-normal cross section of the strand plain and exposed four sandy tsunami deposits, each of which indicates inundation over 2 km inland of the coast. Radiocarbon dating of previously studied and newly discovered deposits in the region indicates a shorter recurrence time for Tokai earthquakes and clarifies their linkage with Nankai earthquakes. We attribute the younger two tsunami deposits to the 1498 and 1096 CE Tokai earthquakes. The older two deposits confirm the occurrence of the Tokai earthquakes in 887 CE and in the latest 7th century. These events are not reliably recorded in historical documents in the Tokai region but were noted in the Nankai area. The 887 CE earthquake likely represents a full-length rupture of the Tokai and Nankai segments, as was the case for the 1707 CE earthquake. Integrated with the previous studies, these new results show that nine Tokai earthquakes occurred over the last 1300 years, the oldest in the latest 7th century, and in 887, 1096, 1361, 1498, 1614, 1707, 1854 and 1944 CE. Recalculated recurrence intervals range from 90 to 265 years. Except for the 1498 Meio Tokai earthquake, the Tokai earthquakes occurred simultaneously with Nankai earthquakes.

Background and objective Amelogenins are major components of extracellular matrix proteins in developing teeth, and regulate the growth of enamel crystals. They also function as signaling molecules in cell differentiation. This study aimed to determine the biological effects of amelogenins on the differentiation of HAT-7 dental epithelial cells and MC3T3-E1 pre-osteoblastic cells using full-length recombinant human amelogenin (rh-AMEL). Design rh-AMEL was expressed in a mammalian cell line (Expi293F™) and was purified by DDK agarose beads. Effects of rh-AMEL on differentiation were evaluated by Mineralization and Alkaline phosphatase (ALP) activity using Alizarin Red S staining and colorimetric substrate p-nitrophenol, respectively. Results Western blotting and silver staining confirmed the successful purification of rh-AMEL. Mineralization and ALP activity in HAT-7 cells were significantly higher after treatment with 4 μg/mL rh-AMEL, but not after treatment with Emdogain ® (EMD). In MC3T3-E1 cells, on the other hand, rh-AMEL showed biphasic effects on differentiation. Treatment with low concentrations of rh-AMEL (0.001–0.1 μg/mL) and EMD (0.01–1 μg/mL) increased mineralization and ALP activity in MC3T3-E1 cells, whereas treatment with high concentrations of rh-AMEL (4 μg/mL) and EMD (100 μg/mL) had the opposite effect. Conclusion High concentrations of rh-AMEL and EMD decreased the differentiation of MC3T3-E1 cells. By contrast, a high concentration of rh-AMEL, but not that of EMD, promoted the differentiation of HAT-7 cells. This study demonstrates that the effects of rh-AMEL on cell differentiation differ between HAT-7 and MC3T3-E1 cells, and suggests that different regions on AMEL may induce the differentiation of these cell types.

Genetically encoded calcium indicators (GECIs) are suitable for long-term imaging studies. In this study, we employed a highly sensitive GECI, G-GECO, and achieved efficient gene delivery with an adenoviral vector. The adenoviral vector allowed us to express G-GECO in more than 80% of cells. More than 80% of G-GECO-expressing cells showed an ATP-induced increase in fluorescence intensity due to Ca

An analytical method using two solid phase extractions and ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was developed for the identification and quantification of 14 mycotoxins (patulin, deoxynivalenol, aflatoxins B1, B2, G1, G2, M1, T-2 toxin, HT-2 toxin, zearalenone, fumonisins B1, B2, B3, and ochratoxin A) in domestic and imported wines. Mycotoxins were purified with an Oasis HLB cartridge, followed by a MultiSepTM #229 Ochra. As a result, sufficient removal of the pigments and highly polar matrices from the red wines was achieved. UHPLC conditions were optimized, and 14 mycotoxins were separated in a total of 13 min. Determinations performed using this method produced high correlation coefficients for the 14 mycotoxins (R > 0.990) and recovery rates ranging from 76 to 105% with good repeatability (relative standard deviation RSD < 12%). Twenty-seven samples of domestic and imported wines were analyzed using this method. Although ochratoxin A (OTA) and fumonisins (FMs) were detected in several samples, the FM levels were less than limits of quantification (LOQs) (1 μg/L), and even the largest of the OTA levels was below the EU regulatory level (2 μg/L). These results suggest that the health risk posed to consumers from the wines available in Japan is relatively low.

Novel polythiophenes with graphene structures (GPTs) could be synthesized based on intramolecular oxidative coupling of the polythiophene precursors having phenyl groups at the 3-position, poly(3-(4′-(3′′,7′′-dimethyloctoxy)phenyl)thiophene) (P3PhT) and poly(3-hexylthiophene)-b-P3PhT (P3HT-b-P3PhT), using FeCl3 as an oxidative agent. The GPTs were characterized by 1H NMR spectroscopy, UV-vis spectroscopy, FT-IR spectroscopy, cyclic voltammetry and X-ray diffraction. The thin film transistor (TFT) characteristics were also performed for P3HT-b-GPT. The progression of oxidative coupling of P3PhTs was confirmed by 1H NMR and FT-IR spectroscopies. Moreover, GPTs showed significant red-shifts in the UV-vis spectra due to their high coplanarity, extended π-conjugation length, and strong π-stacking formation. P3HT-b-GPT also showed a red-shift in the UV-vis spectra and a reduced band gap as compared to the pristine P3HT. P3HT-b-GPT exhibited hole mobility (∼10−5 cm2 V−1 s−1) which is comparable to that of pristine P3HT having a similar molecular weight.

A well-defined diblock copolythiophene, poly(3-hexylthiophene)- b -poly(3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)methylthiophene) (P3HT- b -P3TEGT) was successfully synthesized by a Grignard metathesis (GRIM) polymerization. The structure of the block copolymer was confirmed by size exclusion chromatography (SEC), 1 H NMR spectroscopy, differential scanning calorimetry (DSC), ultraviolet-visible (UV-vis) spectroscopy and fluorescence spectroscopy. The self-assembled structure of the block copolythiophene was investigated by atomic force microscopy (AFM), transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), and synchrotron grazing incidence X-ray scattering (GIXS). These analyses found that the block components in films form vertically oriented lamellar structure via phase separation. Furthermore, both the phases were found to consist of molecular multi-layers respectively, in which the layers stack in the out-of-plane of the film. The P3HT phase exhibited crystalline, which is originated from the π−π stacked thiophene backbones. In contrast, the poly(3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)methylthiophene) (P3TEGT) phase revealed amorphous. Overall, the amphiphilic nature of P3HT- b -P3TEGT successfully demonstrated to lead to well-defined self-assembly structure in films.

Well-defined poly(m-phenylene) (PMP), which is poly(1,3-dibutoxy-m-phenylene), was successfully synthesized via Grignard metathesis polymerization. PMP with a reasonably high number-average molecular weight (Mn) of 25,900 and a very low polydispersity index of 1.07 was obtained. The polymerization of a Grignard reagent monomer, 1-bromo-2,4-dibutoxy-5-chloromagnesiobenzene, proceeded in a chain-growth manner, probably due to the meta-substituted design producing a short distance between the MgCl and Br groups and thereby making a smooth nickel species (CNiC) transfer to the intramolecular chain end (CNiBr) over a benzene ring. PMP showed a good solubility in the common organic solvents, such as tetrahydrofuran, CH2Cl2, and CHCl3. Furthermore, a new block copolymer comprised of PMP and poly(3-hexylthiophene) was also prepared. The tapping mode atomic force microscopy image of the surface of the block copolymer thin film on a mica substrate showed a nanofibril morphology with a clear contrast. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011.

New all-conjugated block copolythiophene, poly(3- hexylthiophene)-block-poly(3-(4 0 -(3 00 ,7 00 -dimethyloctyloxy)-3 0 -pyr- idinyl)thiophene) (P3HT-b-P3PyT) was successfully prepared by Grignard metathesis polymerization. The supramolecular inter- action between (6,6)-phenyl-C61-butyric acid methyl ester (PCBM) and P3PyT was proposed to control the aggregated size of PCBM and long-term thermal stability of the photovol- taic cell, as evidenced by differential scanning calorimetry (DSC), transmission electron microscopy (TEM), and optical mi- croscopy. The effect of different solvents on the electronic and optoelectronic properties was studied, including chloroform (CL), dichlorobenzene (DCB), and mixed solvent of CL/DCB. The optimized bulk heterojunction solar cell devices using the P3HT-b-P3PyT/PCBM blend showed a power conversion effi- ciency of 2.12%, comparable to that of P3HT/PCBM device de- spite the fact that former had a lower crystallinity or absorption coefficient. Furthermore, P3HT-b-P3PyT could be also used as a surfactant to enhance the long-term thermal stability of P3HT/ PCBM-based solar cells by limiting the aggregated size of PCBM. This study represents a new supramolecular approach to design all-conjugated block copolymers for high-performance photovoltaic devices. V C 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 49: 2577-2587, 2011

Regioregular poly(3-(4′-(3′′,7′′-dimethyloctoxy)phenyl)thiophene) (P3PhT) and poly(3-(4′-(3′′,7′′-dimethyloctoxy)-3′-pyridinyl)thiophene) (P3PyT) were successfully prepared with reasonably high mol...

A series of well-defined aniline-chain-end-functionalized regioregular poly(3-hexylthiophene)s (P3HT−NH2) and sulfo-chain-end-functionalized polystyrene (PS−SO3H) have been prepared based on quasi-living Grignard metathesis and living anionic polymerization, respectively. Block copolymers via ionic interaction, (P3HT−NH3+)-b-(PS−SO3−)s, were successfully synthesized, simply by blending P3HT−NH2 with PS−SO3H in toluene. The thermal and optical properties of the block copolymers were investigated by differential scanning calorimetry (DSC) and ultraviolet−visible (UV−vis) spectroscopy. The self-assembly behavior of the (P3HT−NH3+)-b-(PS−SO3−) thin film was observed by atomic force microscopy (AFM) and transmission electron microscopy (TEM). In addition, grazing incidence X-ray scattering (GIXS) analysis found the microphase separation of P3HT−NH3+ and PS−SO3− domains as well as the packing behavior of P3HT−NH3+ segments in block copolymer thin films. By exploiting the pH-sensitive ionic interaction, the PS−S...

The melon 1-aminocyclopropane-1-carboxylic acid oxidase gene ( CM-ACO1 ) is an ethylene responsive gene and plays a role in ethylene synthesis in ripening fruits. To elucidate the regulatory mechanism of the CM-ACO1 gene during the ripening stage, we isolated two cDNAs encoding melon Ethylene Insensitive 3 (EIN3)-like genes ( CmEIL1 and CmEIL2 ) from ripening melon ( Cucumis melo L. cv. Andes) fruits, and studied their properties. The predicted amino acid sequences of CmEIL1 and CmEIL2 were highly homologous to that of EIN3 (60 and 52% similarity for CmEIL1 and CmEIL2, respectively). CmEIL1 and CmEIL2 proteins interacted with a CM-ACO1 promoter in the yeast one-hybrid system, and activated transcription of CM-ACO1 promoter in melon leaf discs by a transient experiment with the luciferase reporter gene. The mRNA levels of CM-ACO1 , CmEIL1 and CmEIL2 genes were very low in immature melon fruits, but increased markedly in fruits during the ripening stage. Ethylene did not affect the expression levels of CmEIL1 and CmEIL2 genes in melon leaf tissues. These results suggest that CmEIL1 and CmEIL2 were ripening-related genes, and that their products acted as transcriptional activators of the CM-ACO1 gene in ripening melon fruits.

A series of well-defined aniline-chain-end-functionalized regioregular poly(3-hexylthiophene)s (P3HT-NH2) have been successfully synthesized on the basis of Grignard metathesis polymerization. The molecular weights of P3HT-NH2 could be varied from 1960 to 28 000 g mol−1, with low polydispersity indices in the range of 1.06–1.29, as determined by gel permeation chromotography. The selectivity of monofunctionalization reactions and the high degrees of amine functionality (85–92%) were confirmed by Matrix-assisted laser desorption ionization time-of-flight mass spectroscopy. Block copolymers through ionic interaction were successfully synthesized by simply blending P3HT-NH2 with carboxy chain-end-functionalized polystyrene, which was prepared on the basis of living anionic polymerization. The thermal and optical properties of block copolymers were investigated by differential scanning calorimetry, Thermogravimetric analysis and UV-vis spectroscopy. The self-assembly behavior of the polymer was investigated by atomic force microscopy and grazing incidence X-ray scattering. Well-defined aniline-chain-end-functionalized regioregular poly(3–hexylthiophene)s (P3HT-NH2) have been synthesized on the basis of Grignard metathesis polymerization. The selectivity of monofunctionalization and the high degree of amine functionality (85–92%) were confirmed by MALDI-TOF mass spectroscopy. A block copolymer through ionic interaction was successfully synthesized simply by blending P3HT-NH2 with carboxy chain-end-functionalized polystyrene. Thermal and optical properties of the block copolymer are investigated by DSC, TGA and UV-vis spectroscopy. The self-assembly behavior of the polymer was observed by AFM showing nanofibril structures with 12–15 nm width.

An HPLC method with a charged aerosol detector (CAD) was optimized and validated for the quantification of six carbohydrates (xylose, fructose, glucose, sucrose, lactose, maltose) and sugar alcohol (erythritol) in drinks. Chromatographic separation was achieved using gradient elution with a mixture of water-methanol (7 : 3) and acetonitril. Carbohydrates were cleaned up using a strong cation exchange cartridge (Bond Elut SCX, Varian). Although an interfering peak appeared without any clean up, it was confirmed that interfering substance had be removed by a treatment using this cartridge and its peak had disappeared. The determinations were performed in the linear range of 5 ∼ 10000 μg/mL for six carbohydrates and sugar alcohol, the correlation coefficients (R) were 0.998 ∼ 1.000, the relative standard deviations (RSDs) ranged over 0.4 ∼ 4.5% (spiked level at 500 μg/mL, n = 5), recoveries ranged between 84 ∼ 113% (spiked level at 500 μg/mL, n = 3), and the limits of quantification were 0.4 ∼ 19.3 μg/mL. In addition, we performed analysis of carbohydrates with CAD, an evaporative light scattering detector (ELSD) and a refractive index detector (RID) to compare and investigate the sensitivity and analysis precision in the each optimum condition. As for the CAD, high sensitivity and good analysis precision were provided. Because it is not necessary to optimize the conditions, CAD is the simplest and easiest device in these detectors. It became clear that CAD was the most suitable to perform correct quantitative analysis of carbohydrates in drinks.