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BACKGROUND: Determining stroke subtypes on initial clinical evaluation is a prerequisite for the selection of appropriate initial treatment. Although diffusion-weighted imaging (DWI) is a powerful tool for detection of acute cerebral infarction, its diagnostic accuracy is not always sufficient particularly in the hyperacute phase. METHODS: Patients admitted within 2 weeks from the symptom onset with the diagnosis of acute ischemic strokes were analyzed with thin-section iso-voxel DWI, namely 3-dimension DWI (3D-DWI), to obtain axial, coronal, and sagittal sections in order to elucidate stroke characteristics. In this case series, we introduce the effectiveness of 3D-DWI. RESULTS: 3D-DWI uncovered stroke subtypes and distribution more precisely compared with conventional DWI. While previous studies indicated the utility of thin section DWI in detecting infratentrial infarctions, 3D-DWI is beneficial for the detection of not only infratentrial but also supratentorial lesions. Furthermore, since both 3D-DWI and magnetic resonance angiography (MRA) are multiplanar reconstruction images, the fusion image of 3D-DWI with MRA is available, enabling cross-reference of spatial cerebrovascular configuration and ischemic lesions. CONCLUSIONS: 3D-DWI is applicable to standard 1.5 T MRI by slight modification of data acquisition protocols, and becomes a key modality to solve the diagnostic puzzle of acute ischemic strokes.
Assuntos
Imagem de Difusão por Ressonância Magnética , Interpretação de Imagem Assistida por Computador , Acidente Vascular Cerebral/diagnóstico por imagem , Idoso , Idoso de 80 Anos ou mais , Angiografia Cerebral , Feminino , Humanos , Angiografia por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Imagem Multimodal , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Acidente Vascular Cerebral/classificação , Acidente Vascular Cerebral/terapiaRESUMO
A 78-year-old man presented with subacute progressive proximal weakness and dysphagia. A biopsy specimen from the left biceps femoris revealed evidence of necrotic and regenerating muscle fibers, but lymphocyte infiltration was not noted. The patient was diagnosed with necrotizing myopathy with anti-signal recognition particle (SRP) antibodies. Concomitant therapy with prednisolone and azathioprine caused the serum CK level to return to normal and it caused clinical manifestations to abate. One year later, however, muscle weakness worsened. Immunoelectrophoresis of serum revealed IgG M protein, and muscle pathology revealed amyloid deposits in numerous blood vessels and at the periphery of a few muscle fibers, and deposits stained positive for anti-λ light chain antibody. The patient was diagnosed with amyloid myopathy, and therapy for systemic amyloid light chain amyloidosis caused muscle weakness to diminish. Amyloidosis is believed to be the primary pathology in this case based on the patient's response to treatment reaction, but the significance of a case involving both amyloid myopathy and necrotizing myopathy warranted examination.
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Amiloidose/diagnóstico , Autoanticorpos/sangue , Doenças Musculares/diagnóstico , Partícula de Reconhecimento de Sinal/imunologia , Idoso , Amiloide/metabolismo , Amiloidose/tratamento farmacológico , Amiloidose/patologia , Biomarcadores/sangue , Biomarcadores/metabolismo , Glicoproteínas/sangue , Humanos , Imunoglobulina G/sangue , Cadeias lambda de Imunoglobulina/metabolismo , Imageamento por Ressonância Magnética , Masculino , Músculos/diagnóstico por imagem , Músculos/metabolismo , Músculos/patologia , Doenças Musculares/tratamento farmacológico , Doenças Musculares/patologia , Necrose , Resultado do TratamentoRESUMO
Osteoporosis is one of the most prevalent ageing-associated diseases that are soaring in the modern world. Although various aspects of the disease have been investigated to understand the bases of osteoporosis, the pathophysiological mechanisms underlying bone loss is still incompletely understood. Poldip2 is a molecule that has been shown to be involved in cell migration of vascular cells and angiogenesis. However, expression of Poldip2 and its regulation in bone cells were not known. Therefore, we examined the Poldip2 mRNA expression and the effects of bone regulators on the Poldip2 expression in osteoblasts. We found that Poldip2 mRNA is expressed in osteoblastic MC3T3-E1 cells. As FGF controls osteoblasts and angiogenesis, FGF regulation was investigated in these cells. FGF suppressed the expression of Poldip2 in MC3T3-E1 cells in a time dependent manner. Protein synthesis inhibitor but not transcription inhibitor reduced the FGF effects on Poldip2 gene expression in MC3T3-E1 cells. As for bone-related hormones, dexamethasone was found to enhance the expression of Poldip2 in osteoblastic MC3T3-E1 cells whereas FGF still suppressed such dexamethasone effects. With respect to function, knockdown of Poldip2 by siRNA suppressed the migration of MC3T3-E1 cells. Poldip2 was also expressed in the primary cultures of osteoblast-enriched cells and FGF also suppressed its expression. Finally, Poldip2 was expressed in femoral bone in vivo and its levels were increased in aged mice compared to young adult mice. These data indicate that Poldip2 is expressed in osteoblastic cells and is one of the targets of FGF. J. Cell. Biochem. 118: 1670-1677, 2017. © 2016 Wiley Periodicals, Inc.
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Fatores de Crescimento de Fibroblastos/farmacologia , Proteínas Mitocondriais/metabolismo , Proteínas Nucleares/metabolismo , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Animais , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Mitocondriais/genética , Proteínas Nucleares/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJECTIVE: The aim of this study was to assess objectively the predictive value of the atypical appearance of the basal layer of oral epithelial dysplasia (OED) for development into invasive carcinoma. STUDY DESIGN: Ninety-six OED cases were examined. These cases were divided into 2 groups: 38 cases that developed into invasive carcinoma and 58 cases that did not. Furthermore, 12 histopathological factors were quantified morphometrically in each case and assessed by Cox's proportional hazards model. RESULTS: The standard deviation of the length between the apical membrane of the basal cells and the basement membrane was significantly associated with development of OED into invasive carcinoma (P < .001; hazard ratio, 3.124). CONCLUSION: We provided novel, objective data demonstrating that an atypical appearance, especially the disordered arrangement of the basal cells representing loss of polarity, may be useful for predicting the development of OED into invasive carcinoma of the tongue.
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Carcinoma Basocelular/patologia , Células Epiteliais/patologia , Mucosa Bucal/patologia , Lesões Pré-Cancerosas/patologia , Neoplasias da Língua/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Membrana Basal/patologia , Biópsia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , PrognósticoRESUMO
Primary tumor (PT) heterogeneity can significantly affect the genetic profile of clones at metastatic sites. To understand the mechanisms underlying metastasis, we compared the genetic profile of paired PT and metastatic lymph node (MLN) samples obtained from patients with oral tongue squamous cell carcinoma (OTSCC). Large-scale genetic profiling was performed on paired PT-MLN samples obtained from 10 OTSCC patients using high-density single-nucleotide polymorphism microarrays. We compared the genetic profile of PT and MLN OTSCC samples to identify common and specific copy number alterations and copy-neutral loss-of-heterozygosity (CN-LOH). Unsupervised hierarchical clustering analysis indicated that 8 of the 10 PT-MLN sample pairs formed clusters, indicating that the primary and metastatic tumors were composed of predominantly genetically similar tumor cells. In 6 of the 10 pairs, 8q11.21, 8q12.2-3, and 8q21.3 gains, and 22q11.23 loss were detected in both the PT and MLN. In addition, 16p11.2 CN-LOH was identified in 9 of the 10 pairs. Conversely, 20q11.2 gain was only observed in the MLNs of 5 of the 10 sample pairs, indicating that genes in this chromosomal region may play a significant role in OTSCC lymph node metastasis. To confirm this, we investigated the expression of two candidate 20q11.2 genes in a separate patient cohort. The expression of one of these genes, E2F1, was significantly increased during the process of metastasis. This study indicates that additional genetic changes, such as 20q11.2 gain, which encodes the E2F1 gene, can be acquired through clonal evolution, and may be required for the metastatic process. © 2016 Wiley Periodicals, Inc.
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Desequilíbrio Alélico/genética , Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Aberrações Cromossômicas , Variações do Número de Cópias de DNA/genética , Neoplasias Bucais/genética , Neoplasias da Língua/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/secundário , Estudos de Casos e Controles , DNA de Neoplasias/genética , Feminino , Seguimentos , Humanos , Perda de Heterozigosidade , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Prognóstico , Neoplasias da Língua/patologiaRESUMO
Bone tissue engineering is in the process of making the shift from bench to bed. Organ as a cell source is important for tissue engineering. The appropriate cells should be harvested without invasiveness and ethical problems. The authors focused on mandibular cortex bone fragments removed in sagittal split ramus osteotomy as a cell source for bone tissue engineering. These bone fragments were discarded after surgery until now. Bone marrow stromal cells (BMSCs) were harvested from inside of bone fragments, which is an endosteal region. Endosteal region is known to be a hematopoietic stem cell niche and harbors osteoblasts, preosteoblasts, and mesenchymal stem cells (MSCs). Bone marrow stromal cells could be cultured easily, and grew rapidly in vitro under ordinary serum-supplemented culture condition. The expression pattern of surface markers of BMSCs was the same as that of MSCs. Bone marrow stromal cells could differentiated into multiple mesenchymal lineages (osteoblasts, adipocytes, chondrocytes, and smooth muscle cells). These results indicated the existence of MSCs in BMSCs. The osteoblastic characters of BMSCs were examined more closely. Bone marrow stromal cells showed a high alkaline phosphatase activity, and expressed osteoblastic markers (PTHr, bone sialoprotein, Type I collagen, Rnut-related transcription factor 2, and osteocalcin). In transplantation experiments, BMSCs generated ectopic bone tissues on the border of hydroxyapatite scaffold without osteogenic differentiation-inducing agents such as dexamethasone (Dex) or bone morphogenetic protein. The results of this study suggest that mandibular cortex bone fragments removed in sagittal split ramus osteotomy are a good cell source for bone tissue engineering.
Assuntos
Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Osteoblastos/citologia , Osteogênese/fisiologia , Osteotomia Sagital do Ramo Mandibular/métodos , Células Estromais/citologia , Engenharia Tecidual/métodos , Diferenciação Celular , Células Cultivadas , Citometria de Fluxo , Humanos , Mandíbula/citologia , Mandíbula/cirurgia , Osteocalcina/metabolismoRESUMO
V domain-containing Ig suppressor of T-cell activation (VISTA)/PD-1H is a novel immune checkpoint molecule for regulating T-cell activation. We examined the effects of anti-VISTA mAb monotherapy and combination therapy with CTLA-4 or PD-1 blockade in a squamous cell carcinoma (SCCVII) model. VISTA monotherapy did not show clear tumor growth regression, but efficiently induced CD8(+) T cell activation by converting resting and exhausted cells into functional effector cells. VISTA monotherapy did not inhibit recruitment of regulatory T cells (Tregs) in the tumor microenvironment (TME). As an additional treatment to VISTA, CTLA-4 blockade, but not PD-1 blockade, elicited further tumor regression. The CTLA-4 and VISTA combination efficiently inhibited Treg recruitment and increased the ratios of both CD8 T/Treg and CD4 conventional T (Tcon)/Treg in the TME, whereas the PD-1 and VISTA combination dramatically increased tumor-recruiting CD8(+) T cells, but markedly reduced the Tcon/Treg ratio. Our results demonstrate that VISTA blockade efficiently converts CD8(+) T cells into functional effector T cells, but is not sufficient to regress tumor growth due to weak Treg suppression in the TME. Our results suggest that combined CTLA-4 and VISTA blockade is more efficacious than combined PD-1 and VISTA blockade for tumors like head and neck squamous cell carcinoma in which Treg-mediated immune regulation is dominant.
Assuntos
Antígenos B7/imunologia , Linfócitos T CD8-Positivos/imunologia , Antígeno CTLA-4/imunologia , Carcinoma de Células Escamosas/imunologia , Receptor de Morte Celular Programada 1/imunologia , Linfócitos T Reguladores/imunologia , Humanos , Ativação Linfocitária , Microambiente TumoralRESUMO
Primary intraosseous squamous cell carcinoma (PIOSCC) is a rare malignant central jaw tumor derived from odontogenic epithelial remnants. PIOSCC predominantly affects the mandible, although both jawbones may be involved. This case report describes a PIOSCC type 2 of the maxilla in a 37-year-old man, treated by partial maxillectomy. Histopathologically, the tumor was diagnosed as PIOSCC derived from an odontogenic cyst. Postoperatively, the patient has been followed up for 53 months, with no recurrence of the disease. We herein describe the clinical details, treatment results and histopathological characteristics of a rare case of PIOSCC derived from a maxillary odontogenic cyst with reference to the relevant literature.
RESUMO
Recent studies have shown that Notch signaling is involved in many types of cancers, including oral squamous cell carcinomas (OSCCs). However, the role of Notch signaling in the tumor microenvironment is not yet fully understood. In this study, we investigated the roles of NOTCH3 signaling in cancer associated fibroblasts (CAFs) in OSCCs. Immunohistochemical study of 93 human tongue OSCC cases indicated that about one third of OSCCs showed NOTCH3 expression in CAFs, and that this expression significantly correlated with tumor-size. In vitro study showed that OSCC cell lines, especially HO1-N-1 cells stimulated NOTCH3 expression in normal human dermal fibroblasts (NHDFs) through direct cell-to-cell contact. Immunohistochemical and morphometric analysis using human OSCC samples demonstrated that NOTCH3 expression in CAFs significantly correlated with micro-vessel density in cancer stroma. In vitro angiogenesis assays involving co-culture of NHDFs with HO1-N-1 and human umbilical endothelial cells (HUVECs), and NOTCH3 knockdown in NHDFs using siRNA, demonstrated that HO1-N-1 cells significantly promoted tube formation dependent on NOTCH3-expression in NHDFs. Moreover, NOTCH3 expression in CAFs was related to poor prognosis of the OSCC patients. This work provides a new insight into the role of Notch signaling in CAFs associated with tumor angiogenesis and the possibility of NOTCH3-targeted molecular therapy in OSCCs.
Assuntos
Carcinoma de Células Escamosas/genética , Fibroblastos/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Bucais/genética , Neovascularização Patológica/genética , Receptor Notch3/genética , Adulto , Idoso , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Comunicação Celular , Técnicas de Cocultura , Feminino , Fibroblastos/patologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/diagnóstico , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Prognóstico , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Receptor Notch3/antagonistas & inibidores , Receptor Notch3/metabolismo , Transdução de Sinais , Análise de Sobrevida , Carga Tumoral , Células Tumorais Cultivadas , Microambiente TumoralRESUMO
INTRODUCTION: Synovial chondromatosis of the temporomandibular joint (TMJ) with cranial extension is rare. Here, we report 7-year follow-up of a case with immunohistochemical examination of cell proliferative activity. PRESENTATION OF CASE: The patient was a 72-year-old man. Severe bone resorption of the glenoid fossa was apparent on CT images. Pathological findings by biopsy led to diagnosis of synovial chondromatosis of the right side TMJ. Extirpation of the tumor was performed via temporopreauricular incision under general anesthesia. PCNA expression was examined by immunohistochemical analysis. The lesion had penetrated into the middle cranial fossa, but the cranial dura mater was intact. Expression of PCNA was confirmed. DISCUSSION: The PCNA expression suggested that growth activity caused expansion of the lesion to the skull base. CONCLUSION: We were able to follow up this case for a long period without recurrence postoperatively.
RESUMO
Statins are recognized as a potential candidate to induce the regeneration of bone. However, statins are a strongly hydrophobic drug and it is difficult to administer at the local sites. In this study, the inclusion complexes of simvastatin (SV) with hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and randomly methylated ß-cyclodextrin (RM-ß-CD) were prepared to improve the water-solubility and the osteogenic differentiation ability of the inclusion complexes in MC3T3-E1 cells was investigated. The water-solubility of SV increased linearly upon the addition of both HP-ß-CD and RM-ß-CD, due to the formation of inclusion complexes. The osteogenic differentiation ability of the inclusion complexes were evaluated by the production of alkaline phosphatase (ALP) and late stage osteogenetic gene expression in MC3T3-E1 cells after 14 days of culture. As a result, the RM-ß-CD/SV inclusion complexes showed significantly higher ALP production and the expression of bone sialoprotein (BSP) and osteocalcin (OCN) than the untreated and free SV-treated cells. Additionally, the production of bone morphogenetic protein-2 (BMP-2) from MC3T3-E1 cells after the treatment with RM-ß-CD/SV inclusion complexes was significantly higher than the untreated and free SV-treated cells. Accordingly, it is concluded that the inclusion complexation of SV with RM-ß-CD is a potential formulation for bone regenerative therapy to improve water-solubility and osteodifferentiation efficiency.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Sinvastatina/química , beta-Ciclodextrinas/química , Fosfatase Alcalina/metabolismo , Animais , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Interações Hidrofóbicas e Hidrofílicas , Camundongos , Osteogênese/efeitos dos fármacos , Sinvastatina/farmacologia , Solubilidade , beta-Ciclodextrinas/farmacologiaRESUMO
OBJECTIVES: Postoperative mandibular fracture (PMF) after marginal resection (MR) of the mandible remains an unresolved issue, and it has been reported that at least 10 mm of postoperative mandibular body height (PMBH) is required to prevent PMF. This study evaluated the clinical, physical, and structural risk factors for PMF in MR patients and determined appropriate preventive measures for PMF. STUDY DESIGN: This retrospective study included 44 patients with lower gingival carcinoma who underwent MR. PMF occurred in four of these patients. Thirteen associated factors identified from medical records and radiographs were statistically analyzed. RESULTS: Mandibular body height (MBH) preservation ratio originally evaluated as less than 0.3, more than 20 remaining teeth after surgery, and inferior alveolar canal (IAC) exposure were significant risk factors for PMF. Prostheses and number of remaining teeth were also correlated with PMF. CONCLUSIONS: The preserved mandibular bone should be reinforced in patients with an MBH preservation ratio of less than 0.3, more than 20 remaining teeth after surgery, and intraoperative IAC exposure. Patients with prostheses are at an increased risk of PMF compared with those without because of stable occlusion and a strong occlusal force. Our novel findings provide useful reference standards for PMF prevention in MR patients.
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Neoplasias Gengivais/cirurgia , Mandíbula/cirurgia , Fraturas Mandibulares/etiologia , Complicações Pós-Operatórias/etiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Neoplasias Gengivais/patologia , Humanos , Arcada Parcialmente Edêntula/diagnóstico por imagem , Masculino , Fraturas Mandibulares/prevenção & controle , Pessoa de Meia-Idade , Gradação de Tumores , Complicações Pós-Operatórias/prevenção & controle , Radiografia Panorâmica , Estudos Retrospectivos , Fatores de RiscoRESUMO
PURPOSE: Bicortical screw fixation systems and miniplate with monocortical screw fixation systems have been reported mainly in bilateral sagittal split ramus osteotomy (BSSO). This study compared postoperative stability between these 2 fixation systems by an intraoral approach. MATERIALS AND METHODS: This was a retrospective cohort study. The study sample was composed of patients treated by BSSO at the authors' institute from January 2006 through December 2012. All cases had facial symmetry and were performed by setback surgery. The predictor variable was treatment group (intraoral screw fixation [SG] vs intraoral miniplate fixation [MG]), and the primary outcome variable was stability defined as the change in the position of point B. Other outcome variables were stability defined as the change in the position of the menton, blood loss, incidence of postoperative temporomandibular joint disorder, and nerve injury. Descriptive and bivariate statistics were computed and the P value was set at .05. RESULTS: Seventy-five patients (35 men and 40 women; mean age, 25.8 yr) were divided into 2 groups (39 SG cases and 36 MG cases). Postoperative changes at point B and the menton in the 2 fixation groups were not statistically different. Lingual nerve injury occurred only in SG cases. Moreover, total blood loss was greater in SG cases. CONCLUSION: An intraoral miniplate with monocortical screw fixation system is recommended over intraoral bicortical screw fixation for bone segments in setback BSSO in patients without facial asymmetry.
Assuntos
Placas Ósseas , Parafusos Ósseos , Osteotomia Sagital do Ramo Mandibular/instrumentação , Adolescente , Adulto , Perda Sanguínea Cirúrgica , Cefalometria/métodos , Queixo/patologia , Estudos de Coortes , Feminino , Seguimentos , Humanos , Traumatismos do Nervo Lingual/etiologia , Masculino , Mandíbula/patologia , Miniaturização , Complicações Pós-Operatórias , Estudos Retrospectivos , Transtornos da Articulação Temporomandibular/etiologia , Resultado do Tratamento , Traumatismos do Nervo Trigêmeo/etiologia , Adulto JovemRESUMO
OBJECTIVES: The aim of the present study was to investigate the factors that contribute to the progression of synovial chondromatosis in the temporomandibular joint (TMJ). METHODS: The authors investigated the expression of CD105 and CD90 in specimens from 17 patients with synovial chondromatosis in the TMJ, using immunohistochemical staining, and expression of CD105 and CD90 in cartilaginous nodules was scored semiquantitatively. RESULTS: The expression of CD105 and CD90 was found in almost all the cases. In particular, the expression of CD90 in cartilaginous nodules significantly decreased with the progression of synovial chondromatosis. DISCUSSION: The factors that determine progression of synovial chondromatosis are not fully understood. The results of this study suggest that CD90 may play an important role in the progression of synovial chondromatosis in the TMJ.
Assuntos
Condromatose Sinovial/metabolismo , Transtornos da Articulação Temporomandibular/metabolismo , Antígenos Thy-1/metabolismo , Adulto , Idoso , Condromatose Sinovial/patologia , Progressão da Doença , Endoglina/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Articulação Temporomandibular/metabolismo , Articulação Temporomandibular/patologia , Transtornos da Articulação Temporomandibular/patologiaRESUMO
Osteoporosis affects over 20 million patients in the United States. Among those, disuse osteoporosis is serious as it is induced by bed-ridden conditions in patients suffering from aging-associated diseases including cardiovascular, neurological, and malignant neoplastic diseases. Although the phenomenon that loss of mechanical stress such as bed-ridden condition reduces bone mass is clear, molecular bases for the disuse osteoporosis are still incompletely understood. In disuse osteoporosis model, bone loss is interfered by inhibitors of sympathetic tone and adrenergic receptors that suppress bone formation. However, how beta adrenergic stimulation affects osteoblastic migration and associated proliferation is not known. Here we introduced a live imaging system, fluorescent ubiquitination-based cell cycle indicator (FUCCI), in osteoblast biology and examined isoproterenol regulation of cell cycle transition and cell migration in osteoblasts. Isoproterenol treatment suppresses the levels of first entry peak of quiescent osteoblastic cells into cell cycle phase by shifting from G1 /G0 to S/G2 /M and also suppresses the levels of second major peak population that enters into S/G2 /M. The isoproterenol regulation of osteoblastic cell cycle transition is associated with isoproterenol suppression on the velocity of migration. This isoproterenol regulation of migration velocity is cell cycle phase specific as it suppresses migration velocity of osteoblasts in G1 phase but not in G1 /S nor in G2 /M phase. Finally, these observations on isoproterenol regulation of osteoblastic migration and cell cycle transition are opposite to the PTH actions in osteoblasts. In summary, we discovered that sympathetic tone regulates osteoblastic migration in association with cell cycle transition by using FUCCI system.
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Osteoblastos/citologia , Osteoblastos/metabolismo , Receptores Adrenérgicos beta/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Pontos de Checagem do Ciclo Celular , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Isoproterenol/farmacologia , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Camundongos , Camundongos Transgênicos , Osteoblastos/efeitos dos fármacos , Hormônio Paratireóideo/farmacologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Célula ÚnicaRESUMO
UNLABELLED: : Although isolation of oral mucosal stromal stem cells has been previously reported, complex isolation methods are not suitable for clinical application. The neurosphere culture technique is a convenient method for the isolation of neural stem cells and neural crest stem cells (NCSCs); neurosphere generation is a phenotype of NCSCs. However, the molecular details underlying the isolation and characterization of human oral mucosa stromal cells (OMSCs) by neurosphere culture are not understood. The purpose of the present study was to isolate NCSCs from oral mucosa using the neurosphere technique and to establish effective in vivo bone tissue regeneration methods. Human OMSCs were isolated from excised human oral mucosa; these cells formed spheres in neurosphere culture conditions. Oral mucosa sphere-forming cells (OMSFCs) were characterized by biological analyses of stem cells. Additionally, composites of OMSFCs and multiporous polylactic acid scaffolds were implanted subcutaneously into immunocompromised mice. OMSFCs had the capacity for self-renewal and expressed neural crest-related markers (e.g., nestin, CD44, slug, snail, and MSX1). Furthermore, upregulated expression of neural crest-related genes (EDNRA, Hes1, and Sox9) was observed in OMSFCs, which are thought to contain an enriched population of neural crest-derived cells. The expression pattern of α2-integrin (CD49b) in OMSFCs also differed from that in OMSCs. Finally, OMSFCs were capable of differentiating into neural crest lineages in vitro and generating ectopic bone tissues even in the subcutaneous region. The results of the present study suggest that OMSFCs are an ideal source of cells for the neural crest lineage and hard tissue regeneration. SIGNIFICANCE: The sphere culture technique is a convenient method for isolating stem cells. However, the isolation and characterization of human oral mucosa stromal cells (OMSCs) using the sphere culture system are not fully understood. The present study describes the isolation of neural crest progenitor cells from oral mucosa using this system. Human OMSCs form spheres that exhibit self-renewal capabilities and multipotency, and are enriched with neural crest-derived cells. These oral mucosa sphere-forming cells can generate ectopic bone tissue in vivo. Therefore, the results of the present study show that the sphere culture system can be applied, without the need for complex isolation techniques, to produce multipotent spheres with the properties of neural crest stem cells. Furthermore, a convenient strategy is demonstrated for the isolation and culture of human OMSCs that could have clinical applications.
Assuntos
Antígenos de Diferenciação/biossíntese , Mucosa Bucal , Células-Tronco Multipotentes , Células-Tronco Neurais , Esferoides Celulares , Adolescente , Adulto , Animais , Células Cultivadas , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/metabolismo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Esferoides Celulares/citologia , Esferoides Celulares/metabolismoRESUMO
We previously reported lower lymphocyte vitamin C levels in individuals with type 2 diabetes mellitus and in individuals with severe Parkinson's disease. Oxidative stress has been proposed to play a key role in the progression of Alzheimer's disease. Thus, the objective of this study was to investigate the association between peripheral levels of vitamin C and the progression of cognitive dysfunction in Alzheimer's disease. Fifty individuals with Alzheimer's disease being treated at Shizuoka General Hospital were consecutively enrolled in this study from December 2009 to March 2015 (76.0±9.7 y of age [mean±SD]; 32 men and 18 women; Mini-Mental State Examination Japanese version (MMSE-J) score range, 8-27). Plasma and lymphocyte vitamin C levels in fasting blood samples were measured. The association between the MMSE-J scores and vitamin C levels was estimated using Spearman's rank correlation coefficient (ρ) and the criteria defined by Swinscow. Spearman's ρ for the relationship between peripheral vitamin C levels and the MMSE-J score was ρ=0.17 for plasma vitamin C and ρ=0.26 for lymphocyte vitamin C. Thus, the associations were relatively weak based on the criteria. In contrast with type 2 diabetes mellitus and Parkinson's disease, lymphocyte vitamin C levels in the peripheral blood may not directly reflect the progression of cognitive dysfunction in Alzheimer's disease. Additional longitudinal studies are needed to evaluate the clinical importance of changes of peripheral vitamin C status in Alzheimer's disease.
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Doença de Alzheimer/sangue , Ácido Ascórbico/sangue , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/prevenção & controle , Ácido Ascórbico/administração & dosagem , Índice de Massa Corporal , Transtornos Cognitivos/tratamento farmacológico , Estudos Transversais , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/efeitos dos fármacosRESUMO
KPU-300 is a novel colchicine-type anti-microtubule agent derived from plinabulin (NPI-2358). We characterized the effects of KPU-300 on cell cycle kinetics and radiosensitization using HeLa cells expressing the fluorescent ubiquitination-based cell cycle indicator (Fucci). Cells treated with 30 nM KPU-300 for 24 h were efficiently synchronized in M phase and contained clearly detectable abnormal Fucci fluorescence. Two-dimensional flow-cytometric analysis revealed a fraction of cells distinct from the normal Fucci fluorescence pattern. Most of these cells were positive for an M phase marker, the phosphorylated form of histone H3. Cells growing in spheroids responded similarly to the drug, and the inner quiescent fraction also responded after recruitment to the growth fraction. When such drug-treated cells were irradiated in monolayer, a remarkable radiosensitization was observed. To determine whether this radiosensitization was truly due to the synchronization in M phase, we compared the radiosensitivity of cells synchronized by KPU-300 treatment and cells in early M phase isolated by a combined method that took advantage of shake-off and the properties of the Fucci system. Following normalization against the surviving fraction of cells treated with KPU-300 alone, the surviving fractions of cells irradiated in early M phase coincided. Taken together with potential vascular disrupting function in vivo, we propose a novel radiosensitizing strategy using KPU-300.
Assuntos
Benzofenonas/farmacologia , Divisão Celular/efeitos dos fármacos , Dicetopiperazinas/farmacologia , Microtúbulos/efeitos dos fármacos , Radiossensibilizantes/farmacologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Citometria de Fluxo , Imunofluorescência , Células HeLa , HumanosRESUMO
INTRODUCTION: Neoplastic disease in the temporomandibular joint (TMJ) is a rare condition and is difficult to differentiate from temporomandibular disorders (TMD) based on symptoms and simple X-ray examinations. Potential differential diagnoses include synovial chondromatosis and pseudotumor, both of which are also relatively rare in the TMJ. PRESENTATION OF CASE: We report a case of pseudotumor of the TMJ that was difficult to differentiate from synovial chondromatosis in a 71-year-old woman with a chief complaint of pain in the left TMJ. MRI of the right TMJ initially led to diagnosis of synovial chondromatosis. Extirpation of the lesion was performed under general anesthesia. Histopathological findings of the resected specimen revealed inflammatory granulation tissue without cellular atypism. DISCUSSION: The pathological findings for the resected specimen were compatible with pseudotumor of the TMJ. These findings were not supportive of synovial chondromatosis or other tumor diseases. CONCLUSION: This case illustrates the importance of careful examination of a mass lesion in the TMJ for differentiation from other TMJ-related diseases.