RESUMO
The placenta plays a fundamental role in fetal growth and maintenance of pregnancy. Its cellular components include a large multinucleated syncytiotrophoblast (STB) and its progenitor, cytotrophoblasts (CTBs), both of which perform vital functions in the human placenta. Primary cytotrophoblasts isolated from term human placentas that spontaneously fuse and differentiate into syncytiotrophoblast-like cells in vitro have been utilized to investigate the functions of the syncytiotrophoblast and placenta with multiple modifications. Although recent advances have enabled the use of trophoblast stem cell-derived organoids as a model for villous trophoblasts, primary CTBs offer several advantages, including spontaneous differentiation, easy access to materials (from term-delivered human placentas), and simple methodology. Here, we present a precise step-by-step process for isolating pure CTBs from term human placenta based on previously reported placenta digestion, density centrifugation, and CTB purification using anti-HLA-A, B, C antibody. Subsequently, we provide a method to improve CTB viability and differentiation into STB-like cells using epidermal growth factor (EGF) and a ROCK inhibitor (Y-27632) that ensures long-term and stable cultures without altering their proliferation. Because these cells can grow on standard tissue culture plates, this model can be easily utilized for various placental investigations, including innate immune responses, drug resistance, and STB metabolism. Employing this approach considerably enhances our understanding of placental functions, which are key to maternal and offspring health.
RESUMO
OBJECTIVE: To clarify whether "low-risk total PP" patients bleed more than partial/marginal PP patients. MATERIALS AND METHODS: The retrospective cohort study was performed involving patients with PP between April 2006 and December 2018. The placental position was determined by ultrasound. From medical charts, the backgrounds as well as obstetric and neonatal outcomes of PP patients were retrieved. RESULTS: This study included 349 patients with PP, which was classified into three types according to the distance between the placenta and internal ostium: total (n = 174), partial (n = 52), and marginal (n = 123) PP. In total PP patients, three factors (prior CS, anterior placenta, and placental lacunae on ultrasound) significantly increased blood loss at CS, the need for hysterectomy, homologous transfusion (≥10 U), and ICU admission. No significant difference was observed in bleeding-related poor outcomes (rate of blood loss ≥2000 mL, amount of homologous transfusion, need for hysterectomy, and ICU admission) between total PP patients without all three factors: "low-risk total PP patients" and partial/marginal PP patients (19.8 vs. 17.1%; p = 0.604, 3.7 vs. 1.1%; p = 0.330, 1.2 vs. 1.1%; p = 1.000, and 1.2 vs. 1.1%; p = 1.000, respectively). CONCLUSION: Prior CS, anterior placenta, and placental lacunae on ultrasound were risk factors for a bleeding-related poor outcome in total PP patients. Total PP patients without these three factors showed the same bleeding-related poor outcome as partial/marginal PP patients.
Assuntos
Placenta Acreta , Placenta Prévia , Feminino , Hemorragia , Humanos , Recém-Nascido , Placenta , Placenta Acreta/terapia , Gravidez , Resultado da Gravidez , Estudos RetrospectivosRESUMO
This study evaluated the usefulness of computed tomography (CT) angiography for the detection of high-risk aortas for carotid artery stenting (CAS). We investigated changes in the treatment method and modifications of endovascular techniques according to the information from CT angiography. CT angiography using three-dimensional and multi-planar reconstruction was performed in 43 consecutive patients with carotid artery stenoses intended to undergo CAS. Two neurosurgeons evaluated the images and estimated the risk and difficulties of CAS. CT angiography clearly depicted the entire aorta, the supra-aortic vessels, and the ilio-femoral arteries in all patients. Abnormal vascular findings were found in 17 patients. High-risk aortic lesions for CAS were detected in 5 patients, including thick aortic plaques with thrombi in 2. We changed the treatment strategy from CAS to carotid endarterectomy (CEA) in these 2 patients. CT angiograms proved very useful in evaluating the approach routes of CAS in 12 patients. One patient had dissecting aneurysm of the thoracic aorta and another had aberrant right vertebral artery. In these two, the treatment strategy was changed from CAS to CEA. The treatment method was changed in 4 patients in total. CT angiography is useful for the detection of high-risk aortas for CAS and for the evaluation of safe approaches to the carotid artery.
Assuntos
Angiografia/instrumentação , Aorta Torácica/diagnóstico por imagem , Doenças da Aorta/diagnóstico , Doenças das Artérias Carótidas/cirurgia , Stents , Adulto , Idoso , Angioplastia/métodos , Doenças da Aorta/complicações , Aterosclerose/complicações , Aterosclerose/diagnóstico , Doenças das Artérias Carótidas/complicações , Doenças das Artérias Carótidas/diagnóstico por imagem , Endarterectomia das Carótidas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Planejamento de Assistência ao Paciente , Tomografia Computadorizada por Raios XRESUMO
A 59-year-old man who was diagnosed with small cell lung cancer (SCLC), achieved a complete response to the induction chemoradiotherapy and received prophylactic cranial irradiation (PCI) (25 Gy at 250 cGy per fraction) in October 2008. Three months later, he complained of anorexia, weight loss, fatigue, and short-term memory loss and developed dementia and systemic muscle weakness. Magnetic resonance imaging in April and July 2009 revealed the progression of the diffuse brain atrophy without evidence of the metastasis of SCLC. Paraneoplastic neurological syndrome was suspected because anti-Hu antibody was detected in his serum and cerebrospinal fluid, but the adverse effects of chemotherapy and/or radiotherapy were also suspected as the cause of his neurological disorder.
Assuntos
Atrofia/etiologia , Encéfalo/patologia , Carcinoma de Células Pequenas/complicações , Demência/etiologia , Neoplasias Pulmonares/complicações , Atrofia/diagnóstico por imagem , Encéfalo/diagnóstico por imagem , Carcinoma de Células Pequenas/diagnóstico por imagem , Carcinoma de Células Pequenas/terapia , Evolução Fatal , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/terapia , Masculino , Pessoa de Meia-Idade , Cintilografia , Terapia de SalvaçãoRESUMO
We present three methods relating to tRNA aminoacylation with non-natural amino acids using an Escherichia coli EF-Tu E215A/D216A mutant that can bind tightly to aa-tRNAs carrying either non-natural or natural amino acids: (i) a method for improving aminoacylation efficiency, (ii) a rapid method for analysing aminoacylation efficiency without the use of radioisotope labelling and (iii) a method for purifying aminoacyl-tRNAs. Although the EF-Tu mutant may be incompatible with some kinds of non-natural amino acids, we confirmed that the EF-Tu mutant could efficiently bind to aa-tRNAs carrying various amino acids (Arg, Ser, O-methyltyrosine, Bodipy FL-aminophenylalanine and 2-acrydonylalanine). These methods may be used for the efficient in vitro synthesis of proteins containing various non-natural amino acids.
Assuntos
Fator Tu de Elongação de Peptídeos/genética , Aminoacil-RNA de Transferência/isolamento & purificação , Aminoacilação de RNA de Transferência/genética , Fator Tu de Elongação de Peptídeos/metabolismoRESUMO
A 40-year-old man presented with a rare case of synchronous multifocal osteosarcoma involving the skull associated with intracerebral hemorrhage, manifesting as sudden headache and left homonymous hemianopia. Computed tomography revealed a skull tumor in the right occipital bone and intracerebral hemorrhage in the right occipital lobe. Gross total resection of the skull tumor with hematoma was performed. The histological diagnosis was osteosarcoma. The tumor cells had invaded into the surrounding brain parenchyma, resulting in intracerebral hemorrhage.
Assuntos
Hemorragia Cerebral/etiologia , Neoplasias Primárias Múltiplas/complicações , Osteossarcoma/patologia , Neoplasias Cranianas/patologia , Adulto , Neoplasias Ósseas/complicações , Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Ósseas/patologia , Hemorragia Cerebral/cirurgia , Evolução Fatal , Humanos , Masculino , Invasividade Neoplásica , Neoplasias Primárias Múltiplas/diagnóstico por imagem , Neoplasias Primárias Múltiplas/patologia , Osteossarcoma/complicações , Osteossarcoma/diagnóstico por imagem , Radiografia , Neoplasias Cranianas/complicações , Neoplasias Cranianas/diagnóstico por imagemRESUMO
A 41-year-old man presented with consciousness disturbance and right hemiparesis. Computed tomography showed a hematoma in the frontal lobe. Left carotid angiography showed a dural arteriovenous fistula (AVF) located in the cranial vault, supplied by the left middle meningeal artery, and drained directly into the cortical vein adjacent to the superior sagittal sinus. Emergency decompressive craniectomy and evacuation of the intracerebral hematoma were performed. A red vein was found on the cortex but the location of the arteriovenous shunt was not clear due to severe brain swelling. Two months later, cranioplasty was performed and a part of the dura mater, expected to have been affected by the dural AVF, was resected. Histological examination disclosed thickening of the intima of the middle meningeal artery and a few small veins around this artery. The fistula was clearly demonstrated between the dural artery and the dural vein. The non-sinal type of dural AVF may originate in similar arteriovenous connections to the sinal type.
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Fístula Arteriovenosa/diagnóstico por imagem , Fístula Arteriovenosa/patologia , Dura-Máter/irrigação sanguínea , Adulto , Humanos , Masculino , RadiografiaRESUMO
This study reports the fabrication and characterization of nanoscale organic light-emitting diodes (nano-OLEDs) based on poly[2-methoxy-5-(2'-ethylhexyloxy)-1,4-phenylene vinylene] (MEH-PPV). The nano-OLEDs were fabricated by spin casting MEH-PPV into cylindrical nanoholes lithographically patterned into silicon nitride. The electroluminescence (EL) spectrum of MEH-PPV was similar to its photoluminescence spectrum, confirming radiative decay from the same excited state. Device characteristics in the form of current density and EL versus applied electric field are presented and compared with those of a large-scale OLED.
Assuntos
Iluminação , Nanoestruturas/química , Nanotecnologia/instrumentação , Compostos Orgânicos/química , Semicondutores , Desenho de Equipamento , Análise de Falha de Equipamento , Teste de Materiais , Nanoestruturas/ultraestrutura , Nanotecnologia/métodosRESUMO
Heparin cofactor II functions as a physiological inhibitor of thrombin activity. The rate of inactivation of thrombin by heparin cofactor II is increased in the presence of dermatan sulfate, which is produced by fibroblasts or smooth muscle cells. To elucidate the role of heparin cofactor II in the extravascular cells, we induced expression of heparin cofactor II in cultured human fibroblasts or vascular smooth muscle cells using adenovirus-mediated gene transfer. After infection of adenovirus vector, these cells secreted heparin cofactor II protein into culture medium. The expressed heparin cofactor II formed the complex with exogenous thrombin and inhibited the proteolytic activity of thrombin. Expression of heparin cofactor II by infection of adenovirus vector inhibited thrombin-induced tissue-type plasminogen activator and interleukin-6 releases from fibroblasts and thrombin-induced interleukin-6 release from vascular smooth muscle cells. These findings show that fibroblasts and vascular smooth muscle cells expressing heparin cofactor II are resistant to thrombin-induced cellular responses.
Assuntos
Adenoviridae/genética , Fibroblastos/efeitos dos fármacos , Cofator II da Heparina/genética , Cofator II da Heparina/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Trombina/antagonistas & inibidores , Células Cultivadas , Cofator II da Heparina/administração & dosagem , Humanos , Interleucina-6/antagonistas & inibidores , Interleucina-6/metabolismo , Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Trombina/farmacologia , Ativador de Plasminogênio Tecidual/antagonistas & inibidores , Ativador de Plasminogênio Tecidual/metabolismo , Transdução GenéticaRESUMO
Calcium spirulan (Ca-SP), a novel sulfated polysaccharide, increases the rate of thrombin inhibition by heparin cofactor II (HCII) more than 1000-fold through a mechanism not requiring the amino-terminal acidic domain of HCII. Activation of HCII by Ca-SP was molecular-weight dependent. Furthermore, HD22, an aptamer that binds exosite II of thrombin, produced a concentration-dependent, 15-fold reduction at 5 microM in the rate of thrombin inhibition by HCII with Ca-SP, suggesting that Ca-SP interacts with exosite II of thrombin. Mutations of Lys173 to Leu (K173L) and Arg189 to Leu (R189L) in the HCII molecule resulted in large decreases in the rate of thrombin inhibition mediated by Ca-SP and in the NaCl concentration needed for elution from Ca-SP-Toyopearl. Mutations of Lys173 to Arg (K173R) and Arg189 to Lys (R189K) showed inhibition of thrombin similar to wild-type rHCII (wt-rHCII). These results indicate that Ca-SP binds to the positive charges of Lys173 and Arg189 on the HCII molecule. In the thrombin inhibitory process by HCII, Ca-SP appears to play as a template by binding to both thrombin and HCII.
Assuntos
Cofator II da Heparina/química , Cofator II da Heparina/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo , Substituição de Aminoácidos , Aminoácidos/química , Sequência de Carboidratos , Cofator II da Heparina/genética , Humanos , Dados de Sequência Molecular , Peso Molecular , Mutagênese Sítio-Dirigida , Trombina/antagonistas & inibidores , Trombina/metabolismoRESUMO
OBJECT: The authors attempted to determine independent predictors that contribute to the recurrence of chronic subdural hematoma (CSDH). METHODS: A total of 105 consecutive patients who underwent surgery for CSDH were included in this study. Eleven patients underwent a repeated operation because the CSDH recurred. Univariate and multivariate analyses were performed to assess the relationships among various variables and CSDH recurrence. Finally, four variables were found to be independently associated with the recurrence of CSDH: 1) absence of a multiplicity of hematoma cavities on CT scans; 2) presence of a history of seizure; 3) width of the hematoma; and 4) absence of a history of diabetes mellitus (DM). CONCLUSIONS: As previously reported, the width of the hematoma is related to the incidence of CSDH recurrence. In this study, the lack of a multiplicity of hematoma cavities was the favorite predictor of CSDH recurrence. In addition, histories of seizure and no past DM are closely related to the incidence of CSDH recurrence.
Assuntos
Hematoma Subdural Crônico/patologia , Hematoma Subdural Crônico/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Seguimentos , Hematoma Subdural Crônico/etiologia , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Procedimentos Neurocirúrgicos/métodos , Valor Preditivo dos Testes , Estudos Prospectivos , Recidiva , Estudos Retrospectivos , Fatores de RiscoRESUMO
The author describes a case of basilar artery occlusion caused by vertebral artery dissection with vertebral fracture. A 61-year-old man was admitted with neck pain after a traffic accident. His symptoms suddenly deteriorated and cerebral angiography revealed an occlusion of the right vertebral artery, and complete occlusion of the basilar artery. Local-arterial fibrinolysis with urokinase for basilar artery occlusion and angioplasty with the use of a self-expandable stent for the site of the vertebral artery dissection was performed, and the basilar artery was partially recanalized. The patient's symptoms gradually improved. It should be emphasized that in cases of acute cervical spine injury after major trauma, vertebral artery dissection should be considered. Local-arterial fibrinolysis and angioplasty using a self-expandable stent was regarded as a useful treatment for basilar artery occlusion caused by vertebral artery dissection, in the acute stage.
Assuntos
Arteriopatias Oclusivas/cirurgia , Artéria Basilar , Vértebras Cervicais , Fraturas da Coluna Vertebral/complicações , Stents , Dissecação da Artéria Vertebral/cirurgia , Fibrinolíticos/uso terapêutico , Heparina/administração & dosagem , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Trombolítica , Procedimentos Cirúrgicos Vasculares/métodos , Dissecação da Artéria Vertebral/etiologiaRESUMO
Most cases of atlas burst fracture do not require surgical stabilization, because they can be successfully treated with external immobilization. The authors present two cases of atlas burst fracture in which surgical stabilization was required after external immobilization. The first patient was a 50-year-old male and the second patient was a 34-year-old male. Both presented with neck pain without neurological symptoms after a traffic accident. Neuroradiological examinations revealed atlas burst fracture in both patients. They were initially treated with conservative treatment; one with a rigid collar and the other with a halo vest. However, lateral offset of the atlas on the axis increased and atlanto-axial instability became evident three months later in both patients. They underwent upper cervical arthrodesis with satisfactory results. The authors review surgical indication and its timing in patients with atlas burst fracture.
Assuntos
Atlas Cervical/lesões , Fraturas da Coluna Vertebral/cirurgia , Adulto , Fixadores Externos , Humanos , Masculino , Pessoa de Meia-Idade , Fusão Vertebral , Resultado do TratamentoRESUMO
A novel strategy for the immobilization of cytochrome c on the surface of chemically modified electrodes is demonstrated and used to investigate the protein's electron-transfer kinetics. Mixed monolayer films of alkanethiols and omega-terminated alkanethiols (terminated with pyridine, imidazole, or nitrile groups that are able to ligate with the heme) are used to adsorb cytochrome c to the surface of gold electrodes. The use of mixed films, as opposed to pure films, allows the concentration of adsorbed cytochrome to remain dilute and ensures a higher degree of homogeneity in their environment. The adsorbed protein is studied using electrochemical methods and scanning tunneling microscopy.
Assuntos
Grupo dos Citocromos c/química , Heme/química , EletroquímicaRESUMO
We evaluated the efficacy of combined surgical and endovascular treatment of high-risk dural arteriovenous fistulas (AVFs) with retrograde leptomeningeal venous drainage. Of 92 patients with intracranial dural AVF, 14 underwent combined surgical and endovascular treatment. Dural AVF was located in the cavernous sinus in two, the petrotentorial region in one, and the transverse-sigmoid sinus in 11. Characteristic features of the patients were retrograde leptomeningeal venous drainage and obstruction of transvenous access routes. All the patients were treated with combined transarterial embolisation and surgical procedures including craniotomy and sinus packing with coils, or clipping of the draining vein. Dural AVF was completely obliterated in all patients without obvious neurologic complications. In the mean follow-up period of 4.2 years, a new dural AVF on the cortex adjacent to the treated sinus developed in one patient. Combined surgical and endovascular treatment is effective for the treatment of high-risk dural AVFs.