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Berlin Brandenburg

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  • Wiley Online Library  (19)
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  • 1
    In: Molecular Microbiology, June 2007, Vol.64(5), pp.1404-1415
    Description: The mechanism of length control of the flagellar hook is under debate between two theories. One claims that the FliK directly measures the hook length as a molecular ruler, while the other claims that the cytoplasmic substructure measures the amount of hook subunits to determine the hook length. Both agree that the FliK C‐terminal domain catalyses the substrate‐specificity switch to terminate hook elongation. In this study, we systematically created mutants with deletions and insertions at various sites within the FliK N‐terminal domain and analysed their effects on the final hook length. Insertions of peptide fragments from the YscP into FliK gave rise to hooks with defined lengths, which was proportional to the molecular size of the FliK‐YscP chimeras. Among deletion mutants, only those with small truncations in three specific sites of FliK produced hooks of a defined, shortened length. For the majority of deletion mutants, FliK was secreted, but hook length was not controlled. On the other hand, for some deletion mutants FliK was not secreted, but the hook length was controlled, indicating that FliK secretion is not necessary for hook‐length control. We conclude that FliK regulates hook length as an internal molecular ruler.
    Keywords: Biology;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 2
    In: FEMS Microbiology Letters, 2006, Vol. 256(1), pp.137-144
    Description: Lipophilic Malassezia species may induce catheter-associated sepsis in premature neonates and immunocompromised patients receiving parenteral lipid emulsions. To assess the participation of lipolytic enzymes in the pathogenesis of this yeast, we cloned a gene encoding the enzyme. A lipolytic enzyme in the culture supernatant of Malassezia pachydermatis was purified 210-fold to homogeneity. The enzyme showed high esterase activity toward p -nitrophenyl octanoate. The cDNA encoding the enzyme was cloned using a degenerate oligonucleotide primer constructed from the N-terminal amino acid sequence. The cDNA consisted of 1582 bp, including an open reading frame encoding 470 amino acids. The first 19 amino acids and the following 13 amino-acid sequence were predicted to be the signal peptides for secretion and prosequence, respectively. The predicted molecular mass of the 438-amino acid mature protein was 48 kDa. Analysis of the deduced amino acid sequence revealed that it contains the consensus motif (Gly–X–Ser–X–Gly), which is conserved among lipolytic enzymes. Homology investigations showed that the enzyme has similarities principally with 11 lipases produced by Candida albicans (29–34% identity) and some other yeast lipases.
    Keywords: 〈Kwd〉〈Italic〉Malassezia Pachydermatis〈/Italic〉〈/Kwd〉 ; Esterase ; Lipase ; Race
    ISSN: 0378-1097
    E-ISSN: 1574-6968
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  • 3
    Language: English
    In: Diabetic Medicine, 10/16/2017
    Description: To purchase or authenticate to the full-text of this article, please visit this link: http://onlinelibrary.wiley.com/doi/10.1111/dme.13524/abstract Byline: K. Iseri, M. Iyoda, Y. Shikida, T. Inokuchi, T. Morikawa, N. Hara, T. Hirano, T. Shibata Abstract Background Type B insulin resistance syndrome is a rare disease characterized by refractory transient hyperglycaemia and severe insulin resistance associated with circulating anti-insulin receptor antibodies. A standardized treatment regimen for type B insulin resistance syndrome has yet to be established. Case report We report the case of a 64-year-old man undergoing haemodialysis for antineutrophil cytoplasmic antibody-associated vasculitis and diabetic nephropathy, who developed rapid onset of hyperglycaemia (glycated albumin 52.1%). Type B insulin resistance syndrome was diagnosed, on the basis of positivity for anti-insulin receptor antibodies and the man's autoimmune history of antineutrophil cytoplasmic antibody-associated vasculitis and idiopathic thrombocytopenic purpura. Although severe hyperglycaemia persisted in spite of corticosteroids and high-dose insulin therapy, rituximab treatment resulted in remarkable improvement of the man's severe insulin resistance and disappearance of anti-insulin receptor antibodies without any adverse effects. Conclusions According to a literature review of 11 cases in addition to the present case, rituximab appears to be a safe and effective strategy for the treatment of corticosteroid-resistant type B insulin resistance syndrome.
    Keywords: Hyperglycemia – Development and Progression ; Hyperglycemia – Care and Treatment ; Insulin Resistance – Development and Progression ; Insulin Resistance – Care and Treatment ; Diabetic Nephropathies – Development and Progression ; Diabetic Nephropathies – Care and Treatment;
    ISSN: Diabetic Medicine
    E-ISSN: 07423071
    E-ISSN: 14645491
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  • 4
    In: Pathology International, April 2016, Vol.66(4), pp.218-223
    Description: Ovarian clear cell carcinomas often show a spherule‐like mucoid stroma. In ascitic fluid, they form spheroids with a hollow acellular space. In spite of the absence of stromal cells, both the mucoid stroma and hollow spheroids contain abundant extracellular matrix, and one of the major components is hyaluronan. It has been suggested that tumor‐derived hyaluronan plays a significant role in the formation of these structures. To clarify this, a hyaluronan inhibition assay was performed on HAC‐2, a clear cell carcinoma cell line, . When hyaluronan synthesis was inhibited by 4‐methylumbelliferone, HAC‐2 failed to show the spherule‐like accumulation of hyaluronan or hollow spheroids. Inhibition of hyaluronan synthesis was associated with the reduction of cell growth. Analysis of 28 archival ascites cytology specimens showed that clear cell carcinomas expressed hyaluronan more frequently than serous carcinomas (11 of 14 3 of 14, respectively, 〈 0.05). All of these facts indicate that tumor‐derived hyaluronan is essential for the formation of the mucoid stroma or hollow spheroids, and that hyaluronan is also involved in the regulation of cell growth in ovarian clear cell carcinomas. The inhibition of hyaluronan synthesis could be a potential adjunctive therapy for refractory clear cell carcinomas outside the ovary.
    Keywords: 4‐Methylumbelliferone ; Ascites ; Clear Cell Carcinoma ; Hollow Spheroid ; Hyaluronan ; Ovary
    ISSN: 1320-5463
    E-ISSN: 1440-1827
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  • 5
    Language: English
    In: Angewandte Chemie International Edition, 04 May 2018, Vol.57(19), pp.5413-5417
    Description: A cationic fluorescent nanogel thermometer based on thermo‐responsive N‐isopropylacrylamide and environment‐sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02–0.84 °C in the range 20–40 °C), and remarkable non‐cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells. thermometer was prepared with a cationic radical initiator. This thermometer is able to enter live mammalian cells within a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02–0.84 °C), and remarkable non‐cytotoxicity, which permitted ordinary cell proliferation and differentiation of primary cultured cells.
    Keywords: Fluorescence Spectroscopy ; Fluorescent Probes ; Imaging Agents ; Nanoparticles ; Sensors
    ISSN: 1433-7851
    E-ISSN: 1521-3773
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  • 6
    In: Genes to Cells, January 2018, Vol.23(1), pp.22-34
    Description: Chronic myeloid leukemia () is caused by the chimeric protein p210 ‐ encoded by a gene on the Philadelphia chromosome. Although the kinase domain of p210 ‐ is an active driver of , the pathological role of its pleckstrin homology () domain remains unclear. Here, we carried out phospholipid vesicle‐binding assays to show that cardiolipin (), a characteristic mitochondrial phospholipid, is a unique ligand of the domain. Arg726, a basic amino acid in the ligand‐binding region, was crucial for ligand recognition. A subset of wild‐type p210 ‐ that was transiently expressed in 293 cells was dramatically translocated from the cytosol to mitochondria in response to carbonyl cyanide ‐chlorophenylhydrazone () treatment, which induces mitochondrial depolarization and subsequent externalization of to the organelle's outer membrane, whereas an R726A mutant of the protein was not translocated. Furthermore, only wild‐type p210 ‐, but not the R726A mutant, suppressed ‐induced mitophagy and subsequently enhanced reactive oxygen species production. Thus, p210 ‐ can change its intracellular localization via interactions between the domain and to cope with mitochondrial damage. This suggests that p210 ‐ could have beneficial effects for cancer proliferation, providing new insight into the domain's contribution to pathogenesis. The chimeric protein p210 ‐ causes chronic myeloid leukemia. We found that cardiolipin is a unique ligand of the domain of p210 ‐. p210 ‐ was dramatically translocated from the cytosol to mitochondria in response to carbonyl cyanide m‐chlorophenylhydrazone () treatment via interactions between the domain and to cope with mitochondrial damage.
    Keywords: Cardiolipin ; Mitochondria ; Mitophagy ; P210 Bcr ‐ Abl ; Pleckstrin Homology Domain ; Reactive Oxygen Species
    ISSN: 1356-9597
    E-ISSN: 1365-2443
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  • 7
    Language: English
    In: Angewandte Chemie, 04 May 2018, Vol.130(19), pp.5511-5515
    Description: A cationic fluorescent nanogel thermometer based on thermo‐responsive N‐isopropylacrylamide and environment‐sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02–0.84 °C in the range 20–40 °C), and remarkable non‐cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells. auf Basis eines fluoreszierenden Nanogels, das mithilfe eines kationischen Radikalinitiators erhalten wurde, kann bei kurzer Inkubationszeit (10 min) in Lebendzellen eindringen. Es reagiert hochempfindlich auf Temperaturveränderungen (Auflösung 0.02–0.84 °C) und ist nicht zellgiftig. Diese Eigenschaften ermöglichten die Proliferation gewöhnlicher Zellen und die Differenzierung von Primärkulturzellen.
    Keywords: Fluoreszenzsonden ; Fluoreszenzspektroskopie ; Kontrastmittel ; Nanopartikel ; Sensoren
    ISSN: 0044-8249
    E-ISSN: 1521-3757
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  • 8
    In: STEM CELLS, February 2016, Vol.34(2), pp.470-482
    Description: Omega‐6 (‐6) and omega‐3 (‐3) polyunsaturated fatty acids (PUFAs) are essential nutrients. Although several studies have suggested that a balanced dietary ‐6:‐3 ratio is essential for brain development, the underlying cellular and molecular mechanism is poorly understood. Here, we found that feeding pregnant mice an ‐6 excess/‐3 deficient diet, which reflects modern human diets, impairsed neocortical neurogenesis in the offspring. This impaired neurodevelopment occurs through a precocious fate transition of neural stem cells from the neurogenic to gliogenic lineage. A comprehensive mediator lipidomics screen revealed key mediators, epoxy metabolites, which were confirmed functionally using a neurosphere assay. Importantly, although the offspring were raised on a well‐balanced ‐6:‐3 diet, they exhibited increased anxiety‐related behavior in adulthood. These findings provide compelling evidence that excess maternal consumption of ‐6 PUFAs combined with insufficient intake of ‐3 PUFAs causes abnormal brain development that can have long‐lasting effects on the offspring's mental state. S C Omega‐6 and omega‐3 polyunsaturated fatty acids are essential nutrients. Although several studies have suggested that a balanced dietary omega‐6:omega‐3 ratio is essential for normal brain development, the underlying cellular and molecular mechanism is poorly understood. Here we shows that maternal consumption of an omega‐6 rich/omega‐3 poor diet impairs offspring's neocortical neuronal layer formation, and reveals its cellular and molecular mechanism; epoxy metabolites of omega‐6 and omega‐3 regulate the fate of neural stem cells (NSCs). We also show that these offspring demonstrate increased anxiety‐related behavior. Our results raise important concerns regarding the increased consumption of such a modern diet, particularly during pregnancy.
    Keywords: Neural Stem Cells ; Arachidonic Acid ; Docosahexaenoic Acid ; Epoxides ; Anxiety ; Metabolomics
    ISSN: 1066-5099
    E-ISSN: 1549-4918
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  • 9
    In: Journal of Neurochemistry, December 2008, Vol.107(5), pp.1248-1260
    Description: Neurotrophins, such as the nerve growth factor (NGF), play an essential role in the growth, development, survival and functional maintenance of neurons in the central and peripheral systems. They also prevent neuronal cell death under various stressful conditions, such as ischemia and neurodegenerative disorders. NGF induces cell differentiation and neurite outgrowth by binding with and activating the NGF receptor tyrosine kinase followed by activation of a variety of signaling cascades. We have investigated the NGF‐dependent neuritogenesis enhancer potential of a food‐derived small molecule contained in vegetables and identified the protein tyrosine phosphatase (PTP) 1B as a key regulator of the NGF receptor‐initiated signal transduction. Based on an extensive screening of vegetable extracts for the neuritogenic‐promoting activity in the rat pheochromocytoma cell line PC12, we found the Japanese horseradish, wasabi (, syn. Eutrema wasabi), as the richest source and identified 6‐methylsulfinylhexyl isothiocyanate (6‐HITC), an analogue of sulforaphane isolated from broccoli, as one of the major neuritogenic enhancers in the wasabi. 6‐HITC strongly enhanced the neurite outgrowth and neurofilament expression elicited by a low‐concentration of NGF that alone was insufficient to induce neuronal differentiation. 6‐HITC also facilitated the sustained‐phosphorylation of the extracellular signal‐regulated kinase and the autophosphorylation of the NGF receptor TrkA. It was found that PTP1B act as a phosphatase capable of dephosphorylating Tyr‐490 of TrkA and was inactivated by 6‐HITC in a redox‐dependent manner. The identification of PTP1B as a regulator of NGF signaling may provide new clues about the chemoprotective potential of food components, such as isothiocyanates.
    Keywords: Isothiocyanate ; Nerve Growth Factor ; Neuritogenesis ; Protein Tyrosine Phosphatase ; Trka
    ISSN: 0022-3042
    E-ISSN: 1471-4159
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  • 10
    In: Acta Crystallographica Section D, November 2004, Vol.60(11), pp.2019-2021
    Description: Regucalcin is a novel calcium ion (Ca) binding protein that does not contain an EF‐hand motif as a Ca‐binding domain and has been demonstrated to play a multi‐functional role in many cell types. Human liver regucalcin, consisting of 299 amino‐acid residues, was overexpressed in , purified and crystallized by the vapour‐diffusion method in the presence of polyethylene glycol 4000 as a precipitant. A native crystal diffracted to 2.8 Å with synchrotron radiation and belongs to space group 2, with unit‐cell parameters  = 64.87, = 52.52, = 86.38 Å, β = 99.86°. Two molecules most probably exist in the asymmetric unit, corresponding to = 2.2 Å Da. Heavy‐atom derivative data were collected and the Pb derivative showed one high‐occupancy site per molecule.
    Keywords: Regucalcin ; Calcium‐Binding Proteins.
    ISSN: 0907-4449
    E-ISSN: 1399-0047
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