A multiple-funnels cell culture insert for the scale-up production of uniform cell spheroids

Introduction: Formation of cell spheres is an important procedure in biomedical research. A large number of high-quality cell spheres of uniform size and shape are required for basic studies and therapeutic applications. Conventional approaches, including the hanging drop method and suspension culture, are used for cell sphere production. However, these methods are time consuming, cell spheres cannot be harvested easily, and it is difficult to control the size and geometry of cell spheres. To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres. Methods: The culture substrate has 680 micro-funnels with a 1-mm width top, 0.89 mm depth, and 0.5 mm square bottom. Mouse embryonic stem cells were used to test the newly developed device. The seeded embryonic stem cells settled at the downward medium surface toward the bottom opening and aggregated as embryoid bodies (EBs). For cell sphere harvest, the bottom of the culture insert was put in contact with the medium surface in another culture dish, and the medium in the device flowed down with cell spheres by hydrostatic pressure. Results: Compact cell spheres with uniform size and shape were collected easily. The diameter of the spheres could be controlled by adjusting the seeding cell density. Spontaneous neural differentiation (nestin and Tju1) and retinoic acid-induced endodermal differentiation (Pdx-1 and insulin I) were improved in the EBs produced using the new insert compared to those in EBs produced by suspension culture. Conclusions: This novel cell culture insert shall improve future studies of cell spheres and benefit clinical applications of cell therapy

Counting Crowds in Bad Weather

Crowd counting has recently attracted significant attention in the field of computer vision due to its wide applications to image understanding. Numerous methods have been proposed and achieved state-of-the-art performance for real-world tasks. However, existing approaches do not perform well under adverse weather such as haze, rain, and snow since the visual appearances of crowds in such scenes are drastically different from those images in clear weather of typical datasets. In this paper, we propose a method for robust crowd counting in adverse weather scenarios. Instead of using a two-stage approach that involves image restoration and crowd counting modules, our model learns effective features and adaptive queries to account for large appearance variations. With these weather queries, the proposed model can learn the weather information according to the degradation of the input image and optimize with the crowd counting module simultaneously. Experimental results show that the proposed algorithm is effective in counting crowds under different weather types on benchmark datasets. The source code and trained models will be made available to the public.Comment: including supplemental materia

LACMA: Language-Aligning Contrastive Learning with Meta-Actions for Embodied Instruction Following

End-to-end Transformers have demonstrated an impressive success rate for Embodied Instruction Following when the environment has been seen in training. However, they tend to struggle when deployed in an unseen environment. This lack of generalizability is due to the agent's insensitivity to subtle changes in natural language instructions. To mitigate this issue, we propose explicitly aligning the agent's hidden states with the instructions via contrastive learning. Nevertheless, the semantic gap between high-level language instructions and the agent's low-level action space remains an obstacle. Therefore, we further introduce a novel concept of meta-actions to bridge the gap. Meta-actions are ubiquitous action patterns that can be parsed from the original action sequence. These patterns represent higher-level semantics that are intuitively aligned closer to the instructions. When meta-actions are applied as additional training signals, the agent generalizes better to unseen environments. Compared to a strong multi-modal Transformer baseline, we achieve a significant 4.5% absolute gain in success rate in unseen environments of ALFRED Embodied Instruction Following. Additional analysis shows that the contrastive objective and meta-actions are complementary in achieving the best results, and the resulting agent better aligns its states with corresponding instructions, making it more suitable for real-world embodied agents. The code is available at: https://github.com/joeyy5588/LACMA.Comment: EMNLP 202

Electrofusion of Mesenchymal Stem Cells and Islet Cells for Diabetes Therapy: A Rat Model

膵島細胞と間葉系幹細胞の融合細胞を用いた糖尿病治療実験に成功 -新しい重症糖尿病治療法の開発に期待-. 京都大学プレスリリース. 2013-05-29.Islet transplantation is a minimally invasive treatment for severe diabetes. However, it often requires multiple donors to accomplish insulin-independence and the long-term results are not yet satisfying. Therefore, novel ways to overcome these problems have been explored. Isolated islets are fragile and susceptible to pro-apoptotic factors and poorly proliferative. In contrast, mesenchymal stem cells (MSCs) are highly proliferative, anti-apoptotic and pluripotent to differentiate toward various cell types, promote angiogenesis and modulate inflammation, thereby studied as an enhancer of islet function and engraftment. Electrofusion is an efficient method of cell fusion and nuclear reprogramming occurs in hybrid cells between different cell types. Therefore, we hypothesized that electrofusion between MSC and islet cells may yield robust islet cells for diabetes therapy. We establish a method of electrofusion between dispersed islet cells and MSCs in rats. The fusion cells maintained glucose-responsive insulin release for 20 days in vitro. Renal subcapsular transplantation of fusion cells prepared from suboptimal islet mass (1, 000 islets) that did not correct hyperglycemia even if co-transplanted with MSCs, caused slow but consistent lowering of blood glucose with significant weight gain within the observation period in streptozotocin-induced diabetic rats. In the fusion cells between rat islet cells and mouse MSCs, RT-PCR showed new expression of both rat MSC-related genes and mouse β-cell-related genes, indicating bidirectional reprogramming of both β-cell and MSCs nuclei. Moreover, decreased caspase3 expression and new expression of Ki-67 in the islet cell nuclei suggested alleviated apoptosis and gain of proliferative capability, respectively. These results show that electrofusion between MSCs and islet cells yield special cells with β-cell function and robustness of MSCs and seems feasible for novel therapeutic strategy for diabetes mellitus

Can machine learning algorithms associated with text mining from internet data improve housing price prediction performance?

Housing frenzies in China have attracted widespread global attention over the past few years, but the key is how to more accurately forecast housing prices in order to establish an effective real estate policy. Based on the ubiquitousness and immediacy of Internet data, this research adopts a broader version of text mining to search for keywords in relation to housing prices and then evaluates the predictive abilities using machine learning algorithms. Our findings indicate that this new method, especially random forest, not only detects turning points, but also offers prediction ability that clearly outperforms traditional regression analysis. Overall, the prediction based on online search data through a machine learning mechanism helps us better understand the trends of house prices in China. First published online 10 June 202

Genome-wide microRNA and gene analysis of mesenchymal stem cell chondrogenesis identifies an essential role and multiple targets for miR-140-5p

microRNAs (miRNAs) are abundantly expressed in development where they are critical determinants of cell differentiation and phenotype. Accordingly miRNAs are essential for normal skeletal development and chondrogenesis in particular. However, the question of which miRNAs are specific to the chondrocyte phenotype has not been fully addressed. Using microarray analysis of miRNA expression during mesenchymal stem cell chondrogenic differentiation and detailed examination of the role of essential differentiation factors, such as SOX9, TGF-b, and the cell condensation phase, we characterize the repertoire of specific miRNAs involved in chondrocyte development, highlighting in particular miR-140 and miR-455. Further with the use of mRNA microarray data we integrate miRNA expression and mRNA expression during chondrogenesis to underline the particular importance of miR-140, especially the -5p strand. We provide a detailed identification and validation of direct targets of miR-140-5p in both chondrogenesis and adult chondrocytes with the use of microarray and 30 UTR analysis. This emphasizes the diverse array of targets and pathways regulated by miR-140-5p. We are also able to confirm previous experimentally identified targets but, additionally, identify a novel positive regulation of the Wnt signaling pathway by miR-140-5p. Wnt signaling has a complex role in chondrogenesis and skeletal development and these findings illustrate a previously unidentified role for miR-140-5p in regulation of Wnt signaling in these processes. Together these developments further highlight the role of miRNAs during chondrogenesis to improve our understanding of chondrocyte development and guide cartilage tissue engineering

Metabolic syndrome and abdominal fat are associated with inflammation, but not with clinical outcomes, in peritoneal dialysis patients

BACKGROUND: In the general population, metabolic syndrome (MetS) is correlated with visceral fat and a risk factor for cardiovascular disease (CVD); however, little is known about the significance of abdominal fat and its association with inflammation and medication use in peritoneal dialysis (PD) patients. We investigated the relationship of visceral fat area (VFA) with C-reactive protein (CRP) levels and medication use in PD patients and followed their clinical outcomes. METHODS: In a prospective study from February 2009 to February 2012, we assessed diabetes mellitus (DM) status, clinical and PD-associated characteristics, medication use, CRP levels, components of MetS, and VFA in 183 PD patients. These patients were categorized into 3 groups based on MetS and DM status: non-MetS (group 1, n = 73), MetS (group 2, n = 65), and DM (group 3, n = 45). VFA was evaluated by computed tomography (CT) and corrected for body mass index (BMI). RESULTS: Patients in group 1 had smaller VFAs than patients in groups 2 and 3 (3.2 ± 1.8, 4.6 ± 1.9, and 4.9 ± 2.0 cm(2)/[kg/m(2)], respectively, P < 0.05) and lower CRP levels (0.97 ± 2.31, 1.27 ± 2.57, and 1.11 ± 1.35 mg/dL, respectively, P < 0.05). VFA increased with the number of criteria met for MetS. After adjusting for age, body weight, and sex, CRP and albumin levels functioned as independent positive predictors of VFA; on other hand, the use of renin-angiotensin system blockers was inversely correlated with VFA in PD patients without DM. In the survival analysis, DM patients (group 3) had the poorest survival among the 3 groups, but no significant differences were found between groups 1 and 2. CONCLUSION: This study showed that VFA and MetS are associated with CRP levels but cannot predict survival in PD patients without DM. The complex relationship of nutritional parameters to VFA and MetS may explain these results. The type of antihypertensive medication used was also associated with the VFA. The mechanisms behind these findings warrant further investigation

AMiBA Wideband Analog Correlator

A wideband analog correlator has been constructed for the Yuan-Tseh Lee Array for Microwave Background Anisotropy. Lag correlators using analog multipliers provide large bandwidth and moderate frequency resolution. Broadband IF distribution, backend signal processing and control are described. Operating conditions for optimum sensitivity and linearity are discussed. From observations, a large effective bandwidth of around 10 GHz has been shown to provide sufficient sensitivity for detecting cosmic microwave background variations.Comment: 28 pages, 23 figures, ApJ in press