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2.
Cell Rep ; 31(1): 107476, 2020 04 07.
Article En | MEDLINE | ID: mdl-32268094

Recent studies using human pluripotent stem cells (hPSCs) have developed protocols to induce kidney-lineage cells and reconstruct kidney organoids. However, the separate generation of metanephric nephron progenitors (NPs), mesonephric NPs, and ureteric bud (UB) cells, which constitute embryonic kidneys, in in vitro differentiation culture systems has not been fully investigated. Here, we create a culture system in which these mesoderm-like cell types and paraxial and lateral plate mesoderm-like cells are separately generated from hPSCs. We recapitulate nephrogenic niches from separately induced metanephric NP-like and UB-like cells, which are subsequently differentiated into glomeruli, renal tubules, and collecting ducts in vitro and further vascularized in vivo. Our selective differentiation protocols should contribute to understanding the mechanisms underlying human kidney development and disease and also supply cell sources for regenerative therapies.


Cell Culture Techniques/methods , Cell Lineage/physiology , Pluripotent Stem Cells/cytology , Cell Differentiation/physiology , Cells, Cultured , Epithelial Cells , Humans , Kidney/cytology , Mesoderm , Nephrons , Organogenesis/physiology , Organoids/cytology , Pluripotent Stem Cells/metabolism , Pluripotent Stem Cells/physiology
9.
J Gen Virol ; 96(8): 2099-2103, 2015 Aug.
Article En | MEDLINE | ID: mdl-25957096

Ticks transmit viruses responsible for severe emerging and re-emerging infectious diseases, some of which have a significant impact on public health. In Japan, little is known about the distribution of tick-borne viruses. In this study, we collected and tested ticks to investigate the distribution of tick-borne arboviruses in Kyoto, Japan, and isolated the first Thogoto virus (THOV) to our knowledge from Haemaphysalis longicornis in far-eastern Asia. The Japanese isolate was genetically distinct from a cluster of other isolates from Africa, Europe and the Middle East. Various cell lines derived from mammals and ticks were susceptible to the isolate, but it was not pathogenic in mice. These results advance understanding of the distribution and ecology of THOV.


Arachnid Vectors/virology , Ixodidae/virology , Thogotovirus/isolation & purification , Tick-Borne Diseases/virology , Animals , Female , Humans , Japan , Male , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Phylogeny , Thogotovirus/classification , Thogotovirus/genetics , Tick-Borne Diseases/transmission
10.
J Virol Methods ; 208: 96-101, 2014 Nov.
Article En | MEDLINE | ID: mdl-25116200

Infection with West Nile virus (WNV), a mosquito-borne flavivirus, is a growing public and animal health concern worldwide. Prevention, diagnosis and treatment strategies for the infection are urgently required. Recently, viral reverse genetic systems have been developed and applied to clinical WNV virology. We developed a protocol for generating reporter virus particles (RVPs) of WNV with the aim of overcoming two major problems associated with conventional protocols, the difficulty in generating RVPs due to the specific skills required for handling RNAs, and the potential for environmental contamination by antibiotic-resistant genes encoded within the genome RNA of the RVPs. By using the proposed protocol, cells were established in which the RVP genome RNA is replicated constitutively and does not encode any antibiotic-resistant genes, and used as the cell supply for RVP genome RNA. Generation of the WNV RVPs requires only the simple transfection of the expression vectors for the viral structural proteins into the cells. Therefore, no RNA handling is required in this protocol. The WNV RVP yield obtained using this protocol was similar that obtained using the conventional protocol. According to these results, the newly developed protocol appears to be a good alternative for the generation of WNV RVPs, particularly for clinical applications.


Genes, Reporter , Molecular Biology/methods , Staining and Labeling/methods , Virology/methods , West Nile virus/physiology , Animals , Cell Line , Cricetinae , Luminescent Proteins/analysis , Luminescent Proteins/genetics , West Nile virus/genetics
11.
Intern Med ; 53(2): 139-43, 2014.
Article En | MEDLINE | ID: mdl-24429455

A 63-year-old man employed in a hard metal manufacturing company for 40 years presented with a chronic dry cough and exertional dyspnea 20 years after the onset of recurrent exanthemas. A chest radiograph revealed bilateral reticular shadows in the upper lung field. Pathological specimens in which tungsten was detected were obtained via a transbronchial lung biopsy. Patch tests were positive for cobalt and other metals. The patient was diagnosed with hard metal lung disease (HMLD) concurrent with contact dermatitis and treated with corticosteroids. This case suggests that allergies to metal may play a role in the onset of HMLD.


Biopsy/methods , Dermatitis, Occupational/etiology , Lung/pathology , Metallurgy , Metals, Heavy/adverse effects , Pneumoconiosis/etiology , Adrenal Cortex Hormones/therapeutic use , Bronchoalveolar Lavage Fluid/cytology , Cobalt/adverse effects , Cobalt/analysis , Dermatitis, Occupational/diagnosis , Dermatitis, Occupational/drug therapy , Electron Probe Microanalysis , Fibrosis , Humans , Immunosuppressive Agents/therapeutic use , Lung/chemistry , Lung/diagnostic imaging , Male , Metals, Heavy/analysis , Middle Aged , Patch Tests , Pneumoconiosis/diagnosis , Pneumoconiosis/diagnostic imaging , Pneumoconiosis/drug therapy , Pneumoconiosis/pathology , Recurrence , Smoking/adverse effects , Tomography, X-Ray Computed , Tungsten/adverse effects , Tungsten/analysis
12.
Intern Med ; 52(23): 2645-51, 2013.
Article En | MEDLINE | ID: mdl-24292756

Fibrosing mediastinitis is rare. One type of this disease is idiopathic fibrosing mediastinitis. It is necessary to rule out malignancy in order to accurately diagnose fibrosing mediastinitis. We herein report a case of anaplastic large cell lymphoma diagnosed three months after a preliminary diagnosis of fibrosing mediastinitis. Glucocorticoid therapy was not successful in controlling disease progression. Immediately after initiating chemotherapy for lymphoma, the patient's symptoms improved dramatically and the mediastinal lesion decreased in size. Although few similar cases have been reported, hidden malignancy may present as fibrosing mediastinitis. Therefore, physicians should consider the probability of malignancy in patients with fibrosing mediastinitis because treatments may vary accordingly.


Lymphoma, Large-Cell, Anaplastic/diagnosis , Mediastinitis/diagnosis , Sclerosis/diagnosis , Adult , Anaplastic Lymphoma Kinase , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Bleomycin/administration & dosage , Cyclophosphamide/administration & dosage , Dacarbazine/administration & dosage , Diagnostic Errors , Doxorubicin/administration & dosage , Female , Fluorodeoxyglucose F18 , Glucocorticoids/therapeutic use , Humans , Lymphoma, Large-Cell, Anaplastic/drug therapy , Mediastinitis/drug therapy , Positron-Emission Tomography , Prednisone/administration & dosage , Radiopharmaceuticals , Receptor Protein-Tyrosine Kinases/metabolism , Sclerosis/drug therapy , Tomography, X-Ray Computed , Vinblastine/administration & dosage , Vincristine/administration & dosage
13.
Kansenshogaku Zasshi ; 87(6): 726-31, 2013 Nov.
Article Ja | MEDLINE | ID: mdl-24483019

Pulmonary Mycobacterium abscessus infection is resistant to many antibiotics and is difficult to treat. We retrospectively analyzed the clinical characteristics of pulmonary infection due to M. abscessus. Eleven cases diagnosed as having pulmonary M. abscessus infection at Osaka Red Cross Hospital from January. 2008, to June, 2012 were enrolled in this study. The average age of the 11 cases was 63 years (all were females). Nine cases showed underlying diseases, comprising 5 cases with Mycobacterium avium complex lung infection, 3 with old pulmonary tuberculosis, and 3 with bronchiectasis. The radiological examination revealed that 10 cases showed the small nodular type, 7 showed the bronchiectatic type, 4 showed a cavity lesion and 4 showed infiltrative shadows. A microbiological definite diagnosis was made from sputum in 10 cases and bronchial lavage fluid in one. As treatment for M. abscessus pulmonary infection, combined multi-drug chemotherapy was carried out in 7 of the 11 cases. No patients were successfully treated with antibiotics alone, whereas 4 patients had no exacerbation of radiological findings without any treatment. One patient received antibiotics including clarithromycin, amikacin and levofloxacin for 2 to 12 months following surgical excision and her sputum cultures have been maintained as negative over the long-term. During the study, none of the 11 patients were known to have died. In this study, we found that M. abscessus pulmonary infection is more common among females, and is found frequently in patients with M. avium complex lung infection. We also found that the clinical course of M. abscessus pulmonary infection was different among patients. We think this is because M. abscessus was shown to comprise three closely related species. M. abscessus is extremely difficult to eradicate, and surgical resection of localized disease or the main lesion or cavity may be significantly effective in preventing the progression of disease.


Lung Diseases , Mycobacterium Infections, Nontuberculous , Nontuberculous Mycobacteria , Adult , Aged , Aged, 80 and over , Female , Humans , Lung Diseases/complications , Lung Diseases/diagnosis , Middle Aged , Mycobacterium Infections, Nontuberculous/complications , Mycobacterium Infections, Nontuberculous/diagnosis , Prospective Studies
14.
Intern Med ; 51(18): 2591-5, 2012.
Article En | MEDLINE | ID: mdl-22989832

Acid-base imbalances and electrolyte disorders induced by proton pump inhibitors (PPIs) are extremely rare. However, under certain conditions, PPIs may cause metabolic acidosis or hypokalemia, probably due to an inhibitory action on the proton pump that contributes to H(+) and K(+) homeostasis in the kidney. We herein present a case of marked hypokalemia accompanied by distal renal tubular acidosis in which a PPI appeared to contribute to the pathophysiology of metabolic acidosis.


Acidosis, Renal Tubular/chemically induced , Gastroesophageal Reflux/drug therapy , Proton Pump Inhibitors/adverse effects , Proton Pump Inhibitors/therapeutic use , Acidosis, Renal Tubular/diagnosis , Female , H(+)-K(+)-Exchanging ATPase/drug effects , Homeostasis/drug effects , Humans , Hydrogen/metabolism , Hypokalemia/chemically induced , Hypokalemia/diagnosis , Kidney/metabolism , Middle Aged , Potassium/metabolism , Proton Pump Inhibitors/pharmacology
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