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  1. Article: Multiple Prostatic Abscesses Caused by Staphylococcus aureus Without Physical Findings in an Immunosuppressed Older Patient.

    Uchiyama, Junji / Tanaka, Yudai / Kurita, Yasuo / Sano, Chiaki / Ohta, Ryuichi

    Cureus

    2023  Volume 15, Issue 1, Page(s) e33555

    Abstract: Staphylococcus ... ...

    Abstract Staphylococcus aureus
    Language English
    Publishing date 2023-01-09
    Publishing country United States
    Document type Case Reports
    ZDB-ID 2747273-5
    ISSN 2168-8184
    ISSN 2168-8184
    DOI 10.7759/cureus.33555
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  2. Article ; Online: Somatostatin affects GnRH neuronal development and migration and stimulates olfactory-related fiber fasciculation.

    Murakami, Shizuko / Ohki-Hamazaki, Hiroko / Uchiyama, Yasuo

    Developmental neurobiology

    2023  Volume 84, Issue 1, Page(s) 3–17

    Abstract: Transient expression of somatostatin (SST) has been observed in the olfactory epithelium (OE) and nerves of chick embryos. Intense expression of SST in these regions on embryonic days (E) 5-8 coincides with the migration of neurons producing gonadotropin- ...

    Abstract Transient expression of somatostatin (SST) has been observed in the olfactory epithelium (OE) and nerves of chick embryos. Intense expression of SST in these regions on embryonic days (E) 5-8 coincides with the migration of neurons producing gonadotropin-releasing hormone (GnRH) from the OE to the forebrain (FB), suggesting that SST plays a role in the development of GnRH neurons. Using in ovo electroporation of small interfering RNA, we found that the suppression of SST mRNA in the olfactory placode (OP) of E3.5 chick embryos significantly reduced the number of GnRH and Islet-1-immunoreactive neurons in the nasal region without affecting the entry of GnRH neurons into the FB at E5.5-6. SST knockdown did not lead to changes in the number of apoptotic, proliferating, or HuC/D-positive neuronal cells in the OE; therefore, it is possible that SST is involved in the neurogenesis/differentiation of GnRH neurons and OP-derived GnRH-negative migratory neurons. In whole OP explant cultures, we also found that SST or its analog octreotide treatment significantly increased the number of migratory GnRH neurons and the migratory distance from the explants. The co-application of an SST antagonist blocked the octreotide-induced increase in the number of GnRH neurons. Furthermore, the fasciculation of polysialylated neural cell adhesion molecule-immunoreactive fibers emerging from the explants was dependent on octreotide. Taken together, our results provide evidence that SST exerts facilitatory effects on the development of neurons expressing GnRH or Islet-1 and on GnRH neuronal migration, in addition to olfactory-related fiber fasciculation.
    MeSH term(s) Animals ; Chick Embryo ; Gonadotropin-Releasing Hormone/metabolism ; Gonadotropin-Releasing Hormone/pharmacology ; Octreotide/metabolism ; Octreotide/pharmacology ; Fasciculation/metabolism ; Neurons/physiology ; Somatostatin/pharmacology ; Somatostatin/metabolism ; Cell Movement/physiology
    Chemical Substances Gonadotropin-Releasing Hormone (33515-09-2) ; Octreotide (RWM8CCW8GP) ; Somatostatin (51110-01-1)
    Language English
    Publishing date 2023-12-10
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2256184-5
    ISSN 1932-846X ; 1097-4695 ; 1932-8451 ; 0022-3034
    ISSN (online) 1932-846X ; 1097-4695
    ISSN 1932-8451 ; 0022-3034
    DOI 10.1002/dneu.22931
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  3. Article: Application of immuno- and affinity labeling with fluorescent dyes to in-resin CLEM of Epon-embedded cells.

    Tanida, Isei / Yamaguchi, Junji / Suzuki, Chigure / Kakuta, Soichiro / Uchiyama, Yasuo

    Heliyon

    2023  Volume 9, Issue 6, Page(s) e17394

    Abstract: In-resin CLEM (Correlative Light and Electron Microscopy) of Epon-embedded cells involves correlating fluorescence microscopy with electron microscopy in the same Epon-embedded ultrathin section. This method offers the advantage of high positional ... ...

    Abstract In-resin CLEM (Correlative Light and Electron Microscopy) of Epon-embedded cells involves correlating fluorescence microscopy with electron microscopy in the same Epon-embedded ultrathin section. This method offers the advantage of high positional accuracy compared to standard CLEM. However, it requires the expression of recombinant proteins. In order to detect the localization of endogenous target(s) and their localized ultrastructures of Epon-embedded samples using in-resin CLEM, we investigated whether immunological and affinity-labeling using fluorescent dyes applied to in-resin CLEM of Epon-embedded cells. The orange fluorescent (λ
    Language English
    Publishing date 2023-06-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 2835763-2
    ISSN 2405-8440
    ISSN 2405-8440
    DOI 10.1016/j.heliyon.2023.e17394
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  4. Article ; Online: Recent advances in in-resin correlative light and electron microscopy of Epon-embedded cells.

    Tanida, Isei / Yamaguchi, Junji / Suzuki, Chigure / Kakuta, Soichiro / Uchiyama, Yasuo

    Microscopy (Oxford, England)

    2023  Volume 72, Issue 5, Page(s) 383–387

    Abstract: Correlative fluorescent and electron microscopic images of the same section of epoxy (or other polymer)-embedded samples, hereafter referred to as 'in-resin CLEM', have been developed to improve the positional accuracy and Z-axis resolution limitations ... ...

    Abstract Correlative fluorescent and electron microscopic images of the same section of epoxy (or other polymer)-embedded samples, hereafter referred to as 'in-resin CLEM', have been developed to improve the positional accuracy and Z-axis resolution limitations of conventional correlative light and electron microscopy (CLEM). High-pressure freezing and quick-freezing substitution result in in-resin CLEM of acrylic-based resin-embedded cells expressing green fluorescent protein, yellow fluorescent protein, mVenus and mCherry, which are sensitive to osmium tetroxide. The identification of osmium-resistant fluorescent proteins leads to the development of in-resin CLEM of Epon-embedded cells. Using subtraction-based fluorescence microscopy with a photoconvertible fluorescent protein, mEosEM-E, its green fluorescence can be observed in thin sections of Epon-embedded cells, and two-color in-resin CLEM using mEosEM-E and mScarlet-H can be performed. Green fluorescent proteins, CoGFP variant 0 and mWasabi, and far-red fluorescent proteins, mCherry2 and mKate2, are available for in-resin CLEM of Epon-embedded cells using the standard procedure for Epon-embedding with additional incubation. Proximity labeling is applied to in-resin CLEM to overcome the limitations of fluorescent proteins in epoxy resin. These approaches will contribute significantly to the future of CLEM analysis.
    MeSH term(s) Humans ; Epoxy Resins ; Microscopy, Electron ; Microscopy, Fluorescence/methods ; Green Fluorescent Proteins ; HeLa Cells
    Chemical Substances Epoxy Resins ; Green Fluorescent Proteins (147336-22-9)
    Language English
    Publishing date 2023-10-03
    Publishing country England
    Document type Journal Article
    ZDB-ID 2707496-1
    ISSN 2050-5701 ; 2050-5698
    ISSN (online) 2050-5701
    ISSN 2050-5698
    DOI 10.1093/jmicro/dfad028
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  5. Article ; Online: TUNEL-positive structures in activated microglia and SQSTM1/p62-positive structures in activated astrocytes in the neurodegenerative brain of a CLN10 mouse model.

    Mitsui, Shun / Yamaguchi, Junji / Suzuki, Chigure / Uchiyama, Yasuo / Tanida, Isei

    Glia

    2023  Volume 71, Issue 12, Page(s) 2753–2769

    Abstract: Neuronal ceroid lipofuscinosis is a group of pediatric neurodegenerative diseases. One of their causative genes, CLN10/CtsD, encodes cathepsin D, a major lysosomal protease. Central nervous system (CNS)-specific CtsD-deficient mice exhibit a ... ...

    Abstract Neuronal ceroid lipofuscinosis is a group of pediatric neurodegenerative diseases. One of their causative genes, CLN10/CtsD, encodes cathepsin D, a major lysosomal protease. Central nervous system (CNS)-specific CtsD-deficient mice exhibit a neurodegenerative disease phenotype with accumulation of ceroid lipofuscins, granular osmiophilic deposits, and SQSTM1/p62. We focused on activated astrocytes and microglia in this neurodegenerative mouse brain, since there are few studies on the relationship between these accumulators and lysosomes in these glial cells. Activated microglia and astrocytes in this mouse thalamus at p24 were increased by approximately 2.5- and 4.6-fold compared with the control, while neurons were decreased by approximately half. Granular osmiophilic deposits were detected in microglial cell bodies and extended their processes in the thalamus. LAMP1-positive lysosomes, but not SQSTM1/p62 aggregates, accumulated in microglia of this mouse thalamus, whereas both lysosomes and SQSTM1/p62 aggregates accumulated in its astrocytes. TUNEL-positive signals were observed mainly in microglia, but few were observed in neurons and astrocytes. These signals were fragmented DNA from degenerated neurons engulfed by microglia or in the lysosomes of microglia. Abnormal autophagic vacuoles also accumulated in the lysosomes of microglia. Granular osmiophilic deposit-like structures localized to LAMP1-positive lysosomes in CtsD-deficient astrocytes. SQSTM1/p62-positive but LAMP1-negative membranous structures also accumulated in the astrocytes and were less condensed than typical granular osmiophilic deposits. These results suggest that CtsD deficiency leads to intracellular abnormalities in activated microglia and astrocytes in addition to neuronal degeneration.
    Language English
    Publishing date 2023-08-12
    Publishing country United States
    Document type Journal Article
    ZDB-ID 639414-0
    ISSN 1098-1136 ; 0894-1491
    ISSN (online) 1098-1136
    ISSN 0894-1491
    DOI 10.1002/glia.24449
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    Zs.A 2345: Show issues Location:
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  6. Article ; Online: Osmium-Resistant Fluorescent Proteins and In-Resin Correlative Light-Electron Microscopy of Epon-Embedded Mammalian Cultured Cells.

    Tanida, Isei / Yamaguchi, Junji / Kakuta, Soichiro / Uchiyama, Yasuo

    Methods in molecular biology (Clifton, N.J.)

    2022  Volume 2564, Page(s) 287–297

    Abstract: Postfixation with osmium tetroxide and Epon embedding are essential for the preservation and visualization of subcellular ultrastructures via electron microscopy. These chemical treatments diminish the fluorescent intensity of most fluorescent proteins ... ...

    Abstract Postfixation with osmium tetroxide and Epon embedding are essential for the preservation and visualization of subcellular ultrastructures via electron microscopy. These chemical treatments diminish the fluorescent intensity of most fluorescent proteins in cells, creating a problem for the in-resin correlative light-electron microscopy (CLEM) of Epon-embedded mammalian cultured cells. We found that two green and two far-red fluorescent proteins retain their fluorescence after chemical fixation with glutaraldehyde, osmium tetroxide-staining, dehydration, and polymerization of Epon resins. Consequently, we could observe the fluorescence of fluorescent proteins in ultrathin sections of Epon-embedded cells via fluorescence microscopy, investigate ultrastructures of the cells in the same sections via electron microscopy, and correlate the fluorescent image with the electron microscopic image without chemical or physical distortion of the cells. In other words, referred as "in-resin CLEM" of Epon-embedded samples. This technique also improves the Z-axis resolution of fluorescent images. In this chapter, we introduce the detailed protocol for in-resin CLEM of Epon-embedded mammalian cultured cells using these fluorescent proteins.
    MeSH term(s) Animals ; Cells, Cultured ; Electrons ; Glutaral ; Mammals ; Microscopy, Electron ; Osmium ; Osmium Tetroxide
    Chemical Substances Osmium (2E7M255OPY) ; Osmium Tetroxide (P40W033BGM) ; Glutaral (T3C89M417N)
    Language English
    Publishing date 2022-09-15
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2667-2_15
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  7. Article ; Online: Wide-ranging migration and destination of early olfactory placode-derived neurons in chick embryos.

    Miyakawa, Momoko / Murakmai, Shizuko / Uchiyama, Yasuo

    Anatomical record (Hoboken, N.J. : 2007)

    2022  Volume 306, Issue 2, Page(s) 298–310

    Abstract: Cell migration from the olfactory placode (OP) is a well-known phenomenon wherein various cell types, such as gonadotropin-releasing hormone (GnRH)-producing neurons, migrate toward the telencephalon (TEL) during early embryonic development. However, the ...

    Abstract Cell migration from the olfactory placode (OP) is a well-known phenomenon wherein various cell types, such as gonadotropin-releasing hormone (GnRH)-producing neurons, migrate toward the telencephalon (TEL) during early embryonic development. However, the spatial relationship between early migratory cells and the forebrain is unclear. We examined the early development of whole-mount chick embryos to observe the three-dimensional spatial relationship among OP-derived migratory neurons, olfactory nerve (ON), and TEL. Migratory neurons that express highly polysialylated neural cell adhesion molecule (PSA-NCAM) emerge from the OP and spread over a relatively wide TEL area at the Hamburger and Hamilton (HH) stage 17. Most migratory neurons form a cellular cord between the olfactory pit and rostral TEL within HH18-20. The earliest axons from the olfactory epithelium (OE) were detected along this neuronal cord using DiI-labeling at HH21. Furthermore, a few PSA-NCAM-positive neurons were dispersed around the cellular cord and over the lateral TEL at HH18. A long cellular cord branch extending to the lateral TEL was transiently observed within HH18-24. These results suggest a novel migratory route of OP-derived neurons during the early developmental stages. Following GFP vector introduction into the OP of HH13-15 embryos, labeled neurons were detected around and within the lateral TEL at HH23 and HH27. At HH36, labeled cells were observed in the rostral-lateral TEL, including the olfactory bulb (OB) region. GFP-labeled and calretinin-positive neurons were detected in the OB, suggesting that early OP-derived neurons enter the forebrain and function as interneurons in the OB.
    MeSH term(s) Animals ; Chick Embryo ; Axons ; Cell Movement ; Neurons/metabolism ; Olfactory Bulb/embryology ; Olfactory Nerve/embryology ; Prosencephalon/embryology ; Telencephalon/embryology
    Language English
    Publishing date 2022-09-22
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2269667-2
    ISSN 1932-8494 ; 1932-8486
    ISSN (online) 1932-8494
    ISSN 1932-8486
    DOI 10.1002/ar.25080
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  8. Article ; Online: Olfactory placode generates a diverse population of neurons expressing GnRH, somatostatin mRNA, neuropeptide Y, or calbindin in the chick forebrain.

    Murakami, Shizuko / Ohki-Hamazaki, Hiroko / Uchiyama, Yasuo

    The Journal of comparative neurology

    2022  Volume 530, Issue 17, Page(s) 2977–2993

    Abstract: The olfactory placode (OP) of vertebrates generates several classes of migrating cells, including hypothalamic gonadotropin-releasing hormone (GnRH)-producing neurons, which play essential roles in the reproduction system. Previous studies using OP cell ... ...

    Abstract The olfactory placode (OP) of vertebrates generates several classes of migrating cells, including hypothalamic gonadotropin-releasing hormone (GnRH)-producing neurons, which play essential roles in the reproduction system. Previous studies using OP cell labeling have demonstrated that OP-derived non-GnRH cells enter the developing forebrain; however, their final fates and phenotypes are less well understood. In chick embryos, a subpopulation of migratory cells from the OP that is distinct from GnRH neurons transiently expresses somatostatin (SS). We postulated that these cells are destined to develop into brain neurons. In this study, we examined the expression pattern of SS mRNA in the olfactory-forebrain region during development, as well as the destination of OP-derived migratory cells, including SS mRNA-expressing cells. Utilizing the Tol2 genomic integration system to induce long-term fluorescent protein expression in OP cells, we found that OP-derived migratory cells labeled at embryonic day (E) 3 resided in the olfactory nerve and medial forebrain at E17-19. A subpopulation of green fluorescent protein (GFP)-labeled GnRH neurons that remained in the olfactory nerve was considered to comprise terminal nerve neurons. In the forebrain, GFP-labeled cells showed a distribution pattern similar to that of GnRH neurons. A large proportion of GFP-labeled cells expressed the mature neuronal marker NeuN. Among the GFP-labeled cells, the percentage of GnRH neurons was low, while the remaining GnRH-negative neurons either expressed SS mRNA, neuropeptide Y, or calbindin D-28k or did not express any of them. These results indicate that a diverse population of OP-derived neuronal cells, other than GnRH neurons, integrates into the chick medial forebrain.
    MeSH term(s) Animals ; Calbindins/metabolism ; Cell Movement/physiology ; Chick Embryo ; Chickens/metabolism ; Gonadotropin-Releasing Hormone/genetics ; Green Fluorescent Proteins/metabolism ; Neurons/metabolism ; Neuropeptide Y/genetics ; Neuropeptide Y/metabolism ; Prosencephalon/metabolism ; RNA, Messenger/metabolism ; Somatostatin/genetics ; Somatostatin/metabolism
    Chemical Substances Calbindins ; Neuropeptide Y ; RNA, Messenger ; Green Fluorescent Proteins (147336-22-9) ; Gonadotropin-Releasing Hormone (33515-09-2) ; Somatostatin (51110-01-1)
    Language English
    Publishing date 2022-07-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3086-7
    ISSN 1096-9861 ; 0021-9967 ; 0092-7317
    ISSN (online) 1096-9861
    ISSN 0021-9967 ; 0092-7317
    DOI 10.1002/cne.25389
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  9. Article: On-Reading (Chinese-Style Pronunciation) Predominance Over Kun-Reading (Native Japanese Pronunciation) in Japanese Semantic Dementia.

    Sakurai, Yasuhisa / Uchiyama, Yumiko / Takeda, Akitoshi / Terao, Yasuo

    Frontiers in human neuroscience

    2021  Volume 15, Page(s) 700181

    Abstract: Japanese kanji (morphograms) have two ways of reading: ...

    Abstract Japanese kanji (morphograms) have two ways of reading:
    Language English
    Publishing date 2021-08-05
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2425477-0
    ISSN 1662-5161
    ISSN 1662-5161
    DOI 10.3389/fnhum.2021.700181
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  10. Article ; Online: Combined 100 keV Cryo-Electron Microscopy and Image Analysis Methods to Characterize the Wider Adeno-Associated Viral Products.

    Nishiumi, Haruka / Hirohata, Kiichi / Fukuhara, Mitsuko / Matsushita, Aoba / Tsunaka, Yasuo / Rocafort, Mark Allen Vergara / Maruno, Takahiro / Torisu, Tetsuo / Uchiyama, Susumu

    Journal of pharmaceutical sciences

    2024  

    Abstract: Adeno-associated viruses (AAVs) are effective vectors for gene therapy. However, AAV drug products are inevitably contaminated with empty particles (EP), which lack a genome, owing to limitations of the purification steps. EP contamination can reduce the ...

    Abstract Adeno-associated viruses (AAVs) are effective vectors for gene therapy. However, AAV drug products are inevitably contaminated with empty particles (EP), which lack a genome, owing to limitations of the purification steps. EP contamination can reduce the transduction efficiency and induce immunogenicity. Therefore, it is important to remove EPs and to determine the ratio of full genome-containing AAV particles to empty particles (F/E ratio). However, most of the existing methods fail to reliably evaluate F/E ratios that are greater than 90 %. In this study, we developed two approaches based on the image analysis of cryo-electron micrographs to determine the F/E ratios of various AAV products. Using our developed convolutional neural network (CNN) and morphological analysis, we successfully calculated the F/E ratios of various AAV products and determined the slight differences in the F/E ratios of highly purified AAV products (purity > 95 %). In addition, the F/E ratios calculated by analyzing more than 1000 AAV particles had good correlations with theoretical F/E ratios. Furthermore, the CNN reliably determined the F/E ratio with a smaller number of AAV particles than morphological analysis. Therefore, combining 100 keV cryo-EM with the developed image analysis methods enables the assessment of a wide range of AAV products.
    Language English
    Publishing date 2024-04-02
    Publishing country United States
    Document type Journal Article
    ZDB-ID 3151-3
    ISSN 1520-6017 ; 0022-3549
    ISSN (online) 1520-6017
    ISSN 0022-3549
    DOI 10.1016/j.xphs.2024.03.026
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