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1.
Sci Rep ; 13(1): 15380, 2023 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-37717055

RESUMO

This paper tackles the challenge of accurate depth estimation from monocular laparoscopic images in dynamic surgical environments. The lack of reliable ground truth due to inconsistencies within these images makes this a complex task. Further complicating the learning process is the presence of noise elements like bleeding and smoke. We propose a model learning framework that uses a generic laparoscopic surgery video dataset for training, aimed at achieving precise monocular depth estimation in dynamic surgical settings. The architecture employs binocular disparity confidence information as a self-supervisory signal, along with the disparity information from a stereo laparoscope. Our method ensures robust learning amidst outliers, influenced by tissue deformation, smoke, and surgical instruments, by utilizing a unique loss function. This function adjusts the selection and weighting of depth data for learning based on their given confidence. We trained the model using the Hamlyn Dataset and verified it with Hamlyn Dataset test data and a static dataset. The results show exceptional generalization performance and efficacy for various scene dynamics, laparoscope types, and surgical sites.


Assuntos
Laparoscopia , Conscientização , Medicamentos Genéricos , Generalização Psicológica , Aprendizado de Máquina Supervisionado
2.
J Exp Biol ; 224(13)2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34170318

RESUMO

Nutrient transfer from mother to embryo is essential for reproduction in viviparous animals. In the viviparous teleost Xenotoca eiseni (family Goodeidae), the intraovarian embryo intakes the maternal component secreted into the ovarian fluid via the trophotaenia. Our previous study reported that the epithelial layer cells of the trophotaenia incorporate a maternal protein via vesicle trafficking. However, the molecules responsible for the absorption were still elusive. Here, we focused on Cubam (Cubilin-Amnionless) as a receptor involved in the absorption, and cathepsin L as a functional protease in the vesicles. Our results indicated that the Cubam receptor is distributed in the apical surface of the trophotaenia epithelium and then is taken into the intracellular vesicles. The trophotaenia possesses acidic organelles in epithelial layer cells and cathepsin L-dependent proteolysis activity. This evidence does not conflict with our hypothesis that receptor-mediated endocytosis and proteolysis play roles in maternal macromolecule absorption via the trophotaenia in viviparous teleosts. Such nutrient absorption involving endocytosis is not a specific trait in viviparous fish. Similar processes have been reported in the larval stage of oviparous fish or the suckling stage of viviparous mammals. Our findings suggest that the viviparous teleost acquired trophotaenia-based viviparity from a modification of the intestinal absorption system common in vertebrates. This is a fundamental study to understand the strategic variation of the reproductive system in vertebrates.


Assuntos
Ciprinodontiformes , Viviparidade não Mamífera , Animais , Endocitose , Feminino , Ovário , Oviparidade
3.
Cell Cycle ; 19(22): 3167-3181, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33131406

RESUMO

During muscle regeneration, extracellular signal-regulated kinase (ERK) promotes both proliferation and migration. However, the relationship between proliferation and migration is poorly understood in this context. To elucidate this complex relationship on a physiological level, we established an intravital imaging system for measuring ERK activity, migration speed, and cell-cycle phases in mouse muscle satellite cell-derived myogenic cells. We found that in vivo, ERK is maximally activated in myogenic cells two days after injury, and this is then followed by increases in cell number and motility. With limited effects of ERK activity on migration on an acute timescale, we hypothesized that ERK increases migration speed in the later phase by promoting cell-cycle progression. Our cell-cycle analysis further revealed that in myogenic cells, ERK activity is critical for G1/S transition, and cells migrate more rapidly in S/G2 phase 3 days after injury. Finally, migration speed of myogenic cells was suppressed after CDK1/2-but not CDK1-inhibitor treatment, demonstrating a critical role of CDK2 in myogenic cell migration. Overall, our study demonstrates that in myogenic cells, the ERK-CDK2 axis promotes not only G1/S transition but also migration, thus providing a novel mechanism for efficient muscle regeneration.


Assuntos
Ciclo Celular/genética , Movimento Celular/genética , Microscopia Intravital/métodos , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/fisiologia , Regeneração/genética , Células Satélites de Músculo Esquelético/metabolismo , Animais , Cardiotoxinas/efeitos adversos , Linhagem Celular , Proliferação de Células/genética , Quinase 2 Dependente de Ciclina/metabolismo , Células Epiteliais/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Humanos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Camundongos Transgênicos , Desenvolvimento Muscular/genética , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/lesões , Transfecção
4.
Development ; 147(9)2020 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-32398354

RESUMO

Osteoblasts arise from bone-surrounding connective tissue containing tenocytes and fibroblasts. Lineages of these cell populations and mechanisms of their differentiation are not well understood. Screening enhancer-trap lines of zebrafish allowed us to identify Ebf3 as a transcription factor marking tenocytes and connective tissue cells in skeletal muscle of embryos. Knockout of Ebf3 in mice had no effect on chondrogenesis but led to sternum ossification defects as a result of defective generation of Runx2+ pre-osteoblasts. Conditional and temporal Ebf3 knockout mice revealed requirements of Ebf3 in the lateral plate mesenchyme cells (LPMs), especially in tendon/muscle connective tissue cells, and a stage-specific Ebf3 requirement at embryonic day 9.5-10.5. Upregulated expression of connective tissue markers, such as Egr1/2 and Osr1, increased number of Islet1+ mesenchyme cells, and downregulation of gene expression of the Runx2 regulator Shox2 in Ebf3-deleted thoracic LPMs suggest crucial roles of Ebf3 in the onset of lateral plate mesoderm differentiation towards osteoblasts forming sternum tissues.


Assuntos
Fatores de Transcrição/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Embrião não Mamífero/metabolismo , Feminino , Fibroblastos/metabolismo , Hibridização In Situ , Proteínas com Homeodomínio LIM/metabolismo , Camundongos , Camundongos Knockout , Osteoblastos/metabolismo , Gravidez , RNA-Seq , Esterno/metabolismo , Fatores de Transcrição/genética , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
5.
J Biophotonics ; 13(5): e201960204, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32078253

RESUMO

Dynamic intravital imaging is essential for revealing ongoing biological phenomena within living organisms and is influenced primarily by several factors: motion artifacts, optical properties and spatial resolution. Conventional imaging quality within a volume, however, is degraded by involuntary movements and trades off between the imaged volume, imaging speed and quality. To balance such trade-offs incurred by two-photon excitation microscopy during intravital imaging, we developed a unique combination of interlaced scanning and a simple image restoration algorithm based on biological signal sparsity and a graph Laplacian matrix. This method increases the scanning speed by a factor of four for a field size of 212 µm × 106 µm × 130 µm, and significantly improves the quality of four-dimensional dynamic volumetric data by preventing irregular artifacts due to the movement observed with conventional methods. Our data suggest this method is robust enough to be applied to multiple types of soft tissue.


Assuntos
Algoritmos , Artefatos , Animais , Microscopia Intravital , Camundongos , Microscopia de Fluorescência , Movimento (Física)
6.
Acta Histochem ; 118(2): 109-17, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26686297

RESUMO

Osteoblast activating peptide (OBAP) was previously reported to be expressed in the rat stomach and to have a vital role in osteogenesis, but its distribution in rat stomach has not been determined. Thus, the aim of the present study was to identify the cell types expressing OBAP in the rat stomach. The stomachs of twelve 10-to-11-week-old male Jc1:SD rats were used. Samples were collected for immunohistochemistry, immunoelectron microscopy and dot blot assay. Immunohistochemical investigation revealed that OBAP was distributed mainly in parietal cells without any expression in chief cells, X/A-like cells or enterochromaffin-like cells. Moreover, OBAP-immunopositive cells were observed mainly in the upper and lower parts of the gastric gland. Significantly high optical density of immunopositive cells was observed in the upper and lower gastric gland regions. The dot blot assay confirmed that OBAP is secreted by parietal cells and that it is present in the gastric gland lumen. Immunoelectron microscopy demonstrated that OBAP was confined to the mitochondrial inner membrane within parietal cells and that the number of mitochondria in the upper and lower parts of the gastric epithelium was significantly larger than the number in the middle part of the gastric epithelium. Based on the results, it was concluded that OBAP is mainly produced by mitochondria of parietal cells in the upper and lower parts of the gastric epithelium. Moreover, the presence of OBAP in the gastric gland lumen suggests an exocrine mechanism of release.


Assuntos
Mucosa Gástrica/metabolismo , Peptídeos/metabolismo , Animais , Masculino , Mitocôndrias/fisiologia , Especificidade de Órgãos , Ratos , Estômago/citologia
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