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关键词： canine   cell culture   corneal endothelial cell   primary cell  

摘要： Objective To establish a primary cell culture and clarify the characteristics of canine corneal endothelial cells in vitro. Procedures The eyes were enucleated from dogs that were euthanized for reasons unrelated to this study. Enucleated canine eyes were dissected, and the intact corneas were isolated from the globes. Using enzymes, the corneal endothelial cells were dispersed from the cornea. The obtained canine corneal endothelial cells were cultured in a cell culture dish. Cultured corneal endothelial cells were morphologically evaluated using phase-contrast microscopy. Immunohistochemical analysis of the cultured cells, particularly of the corneal endothelial cell marker, zonula occludens-1 (ZO-1), Na+/K+-ATPase, and vimentin, was performed to clarify whether the cultured cells were actually corneal endothelial cells. Furthermore, the post-passage morphology of cultured cells was evaluated. Results Canine primary cultured corneal endothelial cells showed morphologically small, cobblestone-like structures. The isolated cells had proliferative ability in vitro and demonstrated positive expression of the corneal endothelial cell markers, ZO-1, Na+/K+-ATPase, and vimentin. However, repeated passages resulted in larger cell sizes as assessed by phase-contrast microscopy. Repeated passages also resulted in lower cell density. Conclusions This study demonstrated the successful culture of canine corneal endothelial cells. This might enhance the understanding of corneal endothelial cell characteristics in dogs.

关键词： Correlative imaging   DNA-PAINT   Live cell imaging   Calcium imaging   Myocytes  

摘要： Nanometre-scale cellular information obtained through super-resolution microscopies are often unaccompanied by functional information, particularly transient and diffusible signals through which life is orchestrated in the nano-micrometre spatial scale. We describe a correlative imaging protocol which allows the ubiquitous intracellular second messenger, calcium (Ca2+), to be directly visualised against nanoscale patterns of the ryanodine receptor (RyR) Ca2+ channels which give rise to these Ca2+ signals in wildtype primary cells. This was achieved by combining total internal reflection fluorescence (TIRF) imaging of the elementary Ca2+ signals, with the subsequent DNA-PAINT imaging of the RyRs. We report a straightforward image analysis protocol of feature extraction and image alignment between correlative datasets and demonstrate how such data can be used to visually identify the ensembles of Ca2+ channels that are locally activated during the genesis of cytoplasmic Ca2+ signals.

关键词： Lentinula edodes   Se-containing polysaccharide   immunosuppressant   T lymphocyte  

摘要： We previously described the biosynthesis, isolation, and immunosuppressive activity of the selenium-containing polysaccharide fraction isolated from the mycelial culture of Lentinula edodes. Structural studies have shown that the fraction was a protein-containing mixture of high molar mass polysaccharides alpha- and beta-glucans. However, which of the components of the complex fraction is responsible for the immunosuppressive activity non-typical for polysaccharides of fungal origin has not been explained. In the current study, we defined four-polysaccharide components of the Se-containing polysaccharide fraction determined their primary structure and examined the effect on T- and B-cell proliferation. The isolated Se-polysaccharides, alpha-1,4-glucan (M-w 2.25 x 10(6) g/mol), unbranched beta-1,6-d-glucan, unbranched beta-1,3-d-glucan and beta-1,3-branched beta-1,6-d-glucan (M-w 1.10 x 10(5) g/mol), are not typical as components of the cell wall of L. edodes. All are biologically active, but the inhibitory effect of the isolated polysaccharides on lymphocyte proliferation was weaker, though more selective than that of the crude fraction.

关键词： Phenotype   Clustering   Cell tracking   Cell morphology   Cell motility  

摘要： In vitro cell culture model systems often employ monocultures, despite the fact that cells generally exist in a diverse, heterogeneous microenvironment in vivo. In response, heterogeneous cultures are increasingly being used to study how cell phenotypes interact. However, the ability to accurately identify and characterize distinct phenotypic subpopulations within heterogeneous systems remains a major challenge. Here, we present the use of a computational, image analysis-based approach-comprising automated contour-based cell tracking for feature identification, principal component analysis for feature reduction, and partitioning around medoids for subpopulation characterization-to non-destructively and non-invasively identify functionally distinct cell phenotypic subpopulations from live-cell microscopy image data. Using a heterogeneous model system of endothelial and smooth muscle cells, we demonstrate that this approach can be applied to both mono and co-culture nuclear morphometric and motility data to discern cell phenotypic subpopulations. Morphometric clustering identified minimal difference in mono- versus co-culture, while motility clustering revealed that a portion of endothelial cells and smooth muscle cells adopt increased motility rates in co-culture that are not observed in monoculture. We anticipate that this approach using non-destructive and non-invasive imaging can be applied broadly to heterogeneous cell culture model systems to advance understanding of how heterogeneity alters cell phenotype.

关键词： Enterprise architecture   theoretical basis   boundary objects theory   cognitive fit theory   management fashion theory   literature review  

摘要： Enterprise architecture is a collection of artifacts describing various aspects of an organization from an integrated business and IT perspective. Practicing enterprise architecture in organizations implies using these artifacts to facilitate information systems planning and improve business and IT alignment. Despite its long history, the enterprise architecture discipline still remains largely atheoretical and lacks a solid theoretical basis. Based on our previous empirical studies of the practical usage of enterprise architecture artifacts in multiple organizations and broad literature analysis, this conceptual article identifies and discusses in detail 10 theories that can be considered key for understanding how an enterprise architecture practice works: actor-network theory, boundary objects theory, cognitive fit theory, communities of practice theory, decision-making theories, information processing theory, knowledge management theory, management fashion theory, media richness theory, and uncertainty principle. Taken together, these theories offer a comprehensive theoretical view of an enterprise architecture practice explaining the role of enterprise architecture artifacts, their usability, and participation of stakeholders and, therefore, may constitute a theoretical basis of the entire enterprise architecture discipline. Although this article does not elaborate on any of these theories, it brings these theories to light, establishes their critical importance for comprehending an enterprise architecture practice, and positions them as central to the enterprise architecture discourse. Each of these theories can be leveraged by enterprise architecture scholars in their future studies for analyzing enterprise architecture practices through respective theoretical lenses. This article intends to provide fresh theoretical insights on enterprise architecture, spark new waves of theoretical enterprise architecture research, and contribute to the development of a soun

关键词： PET   PET/CT   Rituximab   CHOP   Large B-cell lymphoma   Primary mediastinal   Radiotherapy  

摘要： End-of-treatment (EoT) PET/CT is used as a guide to omit radiotherapy (RT) patients with primary mediastinal large B-cell lymphoma (PMBCL). We present the mature and extended results of a retrospective study evaluating the prognostic significance of EoT-PET/CT after adequate response to R-CHOP. Among 231 consecutive PMLBCL patients, 182 underwent EoT-PET/CT and were evaluated according to the Deauville 5-point scale (D5PS) criteria. Freedom from progression (FFP) was measured from the time of PET/CT examination. Among 182 patients, 72 (40%) had D5PS score 1 (D5PSS-1), 33 (18%) had 2, 28 (15%) had 3, 29 (16%) had 4, and 20 (11%) had 5. The 5-year FFP was 97, 94, 92, 82, and 44% for D5PSS-1, D5PSS-2, D5PSS-3, D5PSS-4, and D5PSS-5, respectively. Among 105 patients with unequivocally negative PET/CT (D5PSS-1/D5PSS-2), 49 (47%) received RT (median dose 3420 cGy) and 56 (53%) did not with relapses in 0/49 vs. 4/56 patients (2 mediastinum and 2 isolated CNS relapses).The 5-year FFP for those who received RT or not was 100% versus 96%, when isolated CNS relapses were censored (p = 0.159). Among D5PSS-3 patients (27/28 irradiated-median dose 3600 cGy), the 5-year FFP was 92%. The 5-year FFP for D5PSS-4 and D5PSS-5 was 82 and 44%;44/49 patients received RT (median dose 4000 and 4400 cGy for D5PSS-4 and D5PSS-5). Our study supports the omission of RT in a sizeable fraction of PET/CT-negative patients and definitely discourages salvage chemotherapy and ASCT in patients with PMLBCL who conventionally respond to R-CHOP, solely based on PET/CT positivity in the absence of documented progressive or multifocal disease. The persistence of positive PET/CT with D5PSS < 5 after consolidative RT should not trigger the initiation of further salvage chemotherapy in the absence of conventionally defined PD.

摘要： Take Home MessageAdjuvant therapy with antiangiogenic agents in locoregional renal cell carcinoma did not translate into a sufficient benefit and highlight the crucial role of patient selection. Ongoing clinical trials with immune-targeted agents hold great promise for this unmet clinical need.

关键词： Passive greenhouse systems   glazed balconies   temperature monitoring   multi-family buildings   adaptive comfort  

摘要： Greenhouse systems in the form of glazed balconies may be accomplished both in the newly designed buildings and in the existing ones, raising their energy standard in a quick and inexpensive way. However, basic parameters influencing the efficiency of the systems are often chosen intuitively, not allowing to fully benefit from the sunspaces or causing overheating of the rooms and discomfort for the users. These issues are common drawbacks of passive systems and may become especially important in the aspect of anthropogenic climate changes, including temperature rise and summer heatwaves. The paper presents results of a long-term summer temperature monitoring in flats with glazed balconies of different construction. They were located in prefabricated multi-family buildings, in residential districts of Lublin and Zamosc. The cities are situated in the south-eastern part of Poland, belonging to the warm-summer humid continental climate area. The monitoring enabled overheating assessment according to the concept of adaptive comfort and connecting it with the sunspace construction and the inhabitants' behaviour. These issues are new aspects in the research area, and the work is a part of extensive studies including monitoring and dynamic simulations of dwellings with passive greenhouse systems in Poland.

关键词： Germ cell cancer       Adjuvant chemotherapy       Primary RPLND       Long-term toxicity       Late toxicities  

摘要： Background: One cycle of adjuvant chemotherapy with bleomycin, etoposide and cisplatin (BEP) has shown superiority in recurrence-free survival over retroperitoneal lymph node dissection (RPLND) in patients with clinical stage (CS) I non-seminomatous germ cell tumours (NSGCTs) of the testis in the setting of a phase III trial. We report the recurrences and late toxicities of this study after 13 years of follow-up. Methods: Questionnaires from 382 patients with CS I NSGCT treated with 1 cycle of adjuvant BEP (arm A) or RPLND thorn two cycles of adjuvant BEP in cases of pathological stage II disease (arm B) were evaluated regarding recurrences and late toxicity. Overall, information on recurrence status was available in 337 patients, and 170 questionnaires were evaluable for toxicity (arm A: 95;arm B: 75). Results: With a median follow-up of 13.8 years (0-22), 3 patients (1.6%) in arm A and 16 patients (8.4%) in arm B experienced recurrence. The 15-year PFS in arm A/B was 99% (CI 96-100%)/92% (CI 89-99%) (p = 0.0049). The 15-year OS in arm A/B was 93% (CI 87-97%)/ 93% (CI 86-97%) (p = 0.83). Eight patients (4.2%) in arm A and four patients (2.1%) in arm B showed metachronous secondary testicular cancer (p = 0.26). Five patients (2.6%) in arm A and four patients (2.1%) in arm B developed other malignancies. Toxicities were not significantly different apart from retrograde ejaculation, which occurred more frequently after RPLND (10% versus 24%, p = 0.01). Conclusions: With long-term observation, one cycle of BEP remains superior to RPLND in preventing recurrence and was tolerated without any clinically relevant long-term toxicities. 2021 Elsevier Ltd. All rights reserved.