Density-dependent endothelial cell production of an inhibitor of smooth muscle cell growth

Embryonic data and ultrastructural analyses suggest that the primitive endothelium signals undifferentiated mesenchymal cells to migrate to the forming blood vessel and subsequently regulates mural cell growth and behavior. Upon maturation of the blood vessel, chemotactic and mitogenic signals are apparently diminished and differentiated smooth muscle cells normally remain quiescent. This homeostasis is seemingly upset in conditions which lead to pathologies characterized by smooth muscle cell hyperplasia such as atherosclerosis. By culturing endothelial cells at different densities, we attempted to re-create the various stages of vascular development. Whereas media conditioned by sparse endothelial cells stimulate smooth muscle cells, media conditioned by dense endothelial cell cultures are inhibitory. Culture of sparse smooth muscle cells in media conditioned for 3 days by postconfluent endothelial cell cultures leads to dose-dependent and reversible smooth muscle cell inhibition. Furthermore, in the presence of the endothelial cell-derived inhibitor, smooth muscle cells are rendered refractory to mitogens such as fibroblast growth factor and platelet-derived growth factor. The inhibitory activity is not attributable to the well-characterized inhibitors of smooth muscle cell growth, transforming growth factor type-β, prostaglandin I₂, or heparan sulfate proteoglycan. Partial characterization of the inhibitory conditioned media suggests that the active molecule is smaller than 1,000 da, and stable to boiling as well as proteinase K and heparinase digestion. These findings support the concept that there is intercellular communication between endothelial cells and smooth muscle cells and provide evidence for a novel endothelial cell-derived smooth muscle cell growth inhibitor.     

Deep learning image analysis models pretrained on daily objects are useful for the preliminary characterization of particulate pharmaceutical samples

Visible and subvisible particles are a quality attribute in sterile pharmaceutical samples. A common method for characterizing and quantifying pharmaceutical samples containing particulates is imaging many individual particles using high-throughput instrumentation and analyzing the populations data. The analysis includes conventional metrics such as the particle size distribution but can be more sophisticated by interpreting other visual/morphological features. To avoid the hurdles of building new image analysis models capable of extracting such relevant features from scratch, we propose using well-established pretrained deep learning image analysis models such as EfficientNet. We demonstrate that such models are useful as a prescreening tool for high-level characterization of biopharmaceutical particle image data. We show that although these models are originally trained for completely different tasks (such as the classification of daily objects in the ImageNet database), the visual feature vectors extracted by such models can be useful for studying different types of subvisible particles. This applicability is illustrated through multiple case studies: (i) particle risk assessment in prefilled syringe formulations containing different particle types such as silicone oil, (ii) method comparability with the example of accelerated forced degradation, and (iii) excipient influence on particle morphology with the example of Polysorbate 80 (PS80). As examples of agnostic applicability of pretrained models, we also elucidate the application to two high-throughput microscopy methods: microflow and background membrane imaging. We show that different particle populations with different morphological and visual features can be identified in different samples by leveraging out-of-the-box pretrained models to analyze images from each sample.     

Growth,ROS Markers,Antioxidant Enzymes,Osmotic Regulators and Metabolic Changes in Tartary Buckwheat Subjected to Short Drought

Tartary buckwheat (Fagopyrum tataricum) is an important pseudocereal feed crop with medicinal and nutritional value. Drought is one of the main causes of reduced growth and yield in these plants. We investigated the growth, physiological, and metabolic responses of the widely promoted Tartary buckwheat variety Chuan Qiao No. 1 to polyethylene glycol (PEG)-mediated drought stress. Drought significantly decreased shoot length, shoot biomass and relative water content. Root length, malondialdehyde content, electrolyte leakage, activities of superoxide dismutase, peroxidase, catalase and amylase, and contents of soluble sugar, soluble protein and proline were increased by PEG-mediated drought. Untargeted metabolomics analysis identified 32 core metabolites in seedlings subjected to PEG-mediated drought, 16 of which increased—including quercetin, isovitexin, cyanidin 3-O-beta-D-glucoside, L-arginine, and glycerophosphocholine, while the other 16 decreased—including 3-methoxytyramine, 2, 6-diaminopimelic acid, citric acid, UDP-alpha-D-glucose, adenosine, keto-D-fructose. The 32 core metabolites were enriched in 29 metabolic pathways, including lysine biosynthesis, citrate (TCA) cycle, anthocyanin biosynthesis, and aminoacyl-tRNA biosynthesis. Among them, taurine and hypotaurine metabolism, flavor and flavor biosynthesis, indole alkaline biosynthesis, and alanine, aspartate and glutamate metabolism were the four main metabolic pathways affected by drought. Our findings provide new insights into the physiological and metabolic response mechanisms of Tartary buckwheat to drought stress.     

Identification and expression profiling of receptor-interacting serine/threonine-protein kinase 2 in Siberian sturgeon (Acipenser baerii)

Receptor-interacting serine/threonine-protein kinase 2 (RIPK2) is an adaptor protein of the pattern recognition receptors NOD1 and NOD2 involved in regulating inflammatory response and resisting pathogenic microbial infection. In this study, Acipenser baerii RIPK2 (AbRIPK2) was identified. The open reading frame of AbRIPK2 was 1815 bp encoding 604 amino acids. AbRIPK2 possessed the typical N-terminal kinase domain (KD) and C-terminal caspase recruitment domain (CARD). The phylogenetic tree analysis revealed that AbRIPK2 shared a relatively high identity with bony fish. Real-time fluorescence quantitative PCR (qRT-PCR) results indicated that AbRIPK2 was highly expressed in the gill, followed by muscle, liver and heart. AbRIPK2 was significantly induced in the spleen and valvular intestine after Streptococcus iniae and Aeromonas hydrophila infection. AbRIPK2 was significantly upregulated after peptidoglycan (PGN) treatment in the splenic leukocytes. This study indicated that AbRIPK2 played a potential role in resisting the pathogenic infection of Siberian sturgeon by responding to bacteria.     

低能量激光对造血细胞的增殖作用

激光能刺激造血细胞增殖,促使粒-巨噬细胞集落形成单位(GM—CFUc)增多。从人脐带血中分离制备的有核细胞,加入集落刺激因子(CSF)后,形成的GM—CFUc为18.6±12.6/10~4;如果加入的CSF是经激光照射后的细胞制成的,其GM—CFUc为40.5±20.2/10~4;若先用激光照射脐带血的有核细胞,在培养时加入的是未经激光处理细胞制成的CSF,也有增殖效应,GM—CFUc为36.5±18.5/10~4;如果脐带血有核细胞和制备CSF的细胞都用低能激光照射,就可以见到GM—CFUc明显地增多,其值为57.4±41.2/10~4是未经激光处理者的三倍。表明激光照射脐带血有核细胞及制备CSF的细胞均可使GM—CFUc增殖。     

Identification, sequencing, and targeted mutagenesis of a DNA polymerase gene required for the extreme radioresistance of Deinococcus radiodurans.

Deinococcus radiodurans and other species of the same genus share extreme resistance to ionizing radiation and many other agents that damage DNA. Two different DNA damage-sensitive strains generated by chemical mutagenesis were found to be defective in a gene that has extended DNA and protein sequence homology with polA of Escherichia coli. Both mutant strains lacked DNA polymerase, as measured in activity gels. Transformation of this gene from wild-type D. radiodurans restored to the mutants both polymerase activity and DNA damage resistance. A technique for targeted insertional mutagenesis in D. radiodurans is presented. This technique was employed to construct a pol mutant isogenic with the wild type (the first example of targeted mutagenesis in this eubacterial family). This insertional mutant lacked DNA polymerase activity and was even more sensitive to DNA damage than the mutants derived by chemical mutagenesis. In the case of ionizing radiation, the survival of the wild type after receiving 1 Mrad was 100% while survival of the insertional mutant extrapolated to 10(-24). These results demonstrate that the gene described here encodes a DNA polymerase and that defects in this pol gene cause a dramatic loss of resistance of D. radiodurans to DNA damage.     

Injecting skin fibroblasts into muscles to express human clotting factor IX cDNA

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球孢白僵菌继代培养中菌落局变现象及环境影响因素的研究

球孢白僵菌[Beauveria bassiana(Bals.)Vuill.]野生型多孢株以及从其上分离的单孢株在不同培养基、光照、温度和湿度条件下继代培养,结果显示:无沦是多孢株还是单孢株,在传代中均发生显著的菌落局变,并产生形态各异的分离子,多表现为菌苔瘠薄或气生菌丝徒长,肉眼可见产孢量减少;尤其是单孢株菌落局变产生的分离子往往伴随着棕黄色或红棕色色素的分泌。培养基、温度、湿度和光照等环境因素对菌落局变产生具明显诱导作用。以SDAY培养基、偏低的温湿度和全光照条件下产生局变频率较低。大批量生产中以麦麸作原料较适宜。提出了菌落局变发生的几种可能的遗传机制。     

扫描电镜分析牙齿釉质结构方法的讨论

通过扫描电镜对大熊猫牙齿釉质结构的三维观察与比较,结果表明:在同一枚牙齿上,不同部位的釉质切面,釉柱截面的形状、大小以及排列等,有不同的现象.因此在用上述特征描述时,应对所有要比较的样品,须精确地统一在相同的部位.本文以多次实验结果,表明了严格定位取样的重要性.     

云南引种的玫瑰天竺葵精油主要化学成份分析

玫瑰天竺葵从摩洛哥引入云南已试种成功,并开始大面积推广。其各生长期精油分析结果表明,随月份的推移,香叶醇含量下降。其香气特征与云南产香叶天竺葵(Pelargonium graveolens)精油不同,更接近印度波尔蓬香叶油的香气。