High Density Culture Process and Growth Kinetics of Flavor Yeast A10-2

In order to realize the high-density culture and establish the predictive growth kinetics model of a self-isolated flavor yeast, the high-density culture process was studied of 4-ethyl-2-methoxyphenol producing yeast A10-2, which screened from soy sauce fermentation mash. The type of culture medium (nitrogen and carbon source) and concentration were studied and optimized. The growth kinetics and the substrate (total sugar) consumption models were established and verified. The results showed that (NH4) H2PO4 fed to make the concentration of culture medium 0.2 g/100 mL was the best inorganic source of nitrogen. To obtain the best cell growth rate, molasses as the only source of carbon, should be fed which controlled the concentration of total sugar in the culture medium to 0.4~0.6 g/100 mL. The growth of A10-2 yeast followed the S-shaped curve of a Logistic model, and the substrate (total sugar) consumption followed the Leudeking-Priet equation. The maximum obtained biomass specific growth rate μm was 0.4764 h−1, while the maximum biomass growth yield coefficient YG was 0.5879 g/g. The maintenance coefficient was 0.0127 g·L−1·h−1. The established models could better describe the growth and sugar consumption of yeast in the process of high-density culture, and have predictive significance

Variations of <i>CITED2</i> Are Associated with Congenital Heart Disease (CHD) in Chinese Population

<div><p><i>CITED2</i> was identified as a cardiac transcription factor which is essential to the heart development. <i>Cited2</i>-deficient mice showed cardiac malformations, adrenal agenesis and neural crest defects. To explore the potential impact of mutations in <i>CITED2</i> on congenital heart disease (CHD) in humans, we screened the coding region of <i>CITED2</i> in a total of 700 Chinese people with congenital heart disease and 250 healthy individuals as controls. We found five potential disease-causing mutations, p.P140S, p.S183L, p.S196G, p.Ser161delAGC and p. Ser192_Gly193delAGCGGC. Two mammalian two-hybrid assays showed that the last four mutations significantly affected the interaction between <i>p300CH1</i> and <i>CITED2</i> or <i>HIF1A</i>. Further studies showed that four <i>CITED2</i> mutations recovered the promoter activity of <i>VEGF</i> by decreasing its competitiveness with <i>HIF1A</i> for binding to <i>p300CH1</i> and three mutations decreased the consociation of <i>TFAP2C</i> and <i>CITED2</i> in the transactivation of <i>PITX2C</i>. Both <i>VEGF</i> and <i>PITX2C</i> play very important roles in cardiac development. In conclusion, we demonstrated that <i>CITED2</i> has a potential causative impact on congenital heart disease.</p></div

Effect of <i>CITED2</i> variants on the cooperation between <i>CITED2</i> and <i>TFAP2C</i> in the transactivation of the <i>PITX2C</i>.

<p><b>A:</b> Effect of <i>CITED2</i> mutations on the transcription activation of <i>PITX2C</i>. (*p<0.05, **p<0.01 versus wt-type, #p<0.05, ##p<0.01 versus.empty vector pcDNA3.1(+)). <b>B: </b><i>CITED2</i>-wt and <i>TFAP2C</i> working on the transcriptional activation of <i>PITX2C</i>. <i>PITX2C</i> reporter plasmid and the expression vector for <i>TFAP2C</i>, <i>CITED2</i>, or pcDNA3.1 alone were transfected respectively in 293 T cells. The luciferase activity was normalized to Renilla activity.(* p<0.05, **p<0.01 versus the untreated group (n = 3)).</p

Primers used for PCR.

<p>Primers used for PCR.</p

Interfacial Nanoinjection-Based Nanoliter Single-Cell Analysis

Single-cell analysis offers unprecedented resolution for the investigation of cellular heterogeneity and the capture of rare cells from large populations. Here, described is a simple method named interfacial nanoinjection (INJ), which can miniaturize various single-cell assays to be performed in nanoliter water-in-oil droplets on standard microwell plates. The INJ droplet handler can adjust droplet volumes for multistep reactions on demand with high precision and excellent monodispersity, and consequently enables a wide range of single-cell assays. Importantly, INJ can be coupled with fluorescence-activated cell sorting (FACS), which is currently the most effective and accurate single-cell sorting and isolation method. FACS-INJ pipelines for high-throughput plate well-based single-cell analyses, including single-cell proliferation, drug-resistance testing, polymerase chain reaction (PCR), reverse-transcription PCR, and whole-genome sequencing are introduced. This FACS-INJ pipeline is compatible with a wide range of samples and can be extended to various single-cell analysis applications in microbiology, cell biology, and biomedical diagnostics

Patients with congenital heart disease included in the study.

<p>Patients with congenital heart disease included in the study.</p

Structure of <i>CITED2</i>.

<p><b>A</b>: Sequence alignment of <i>CITED2</i> proteins among several species. The figure showed that three acid substitutions were located at highly conserved regions among many species (human, chimpanzee, mice, dog, cattle, rat, chicken and zebrafish). <b>B</b>: Position of mutations in the <i>CITED2</i> protein identified in CHD patients. <i>CITED2</i> has three conserved regions CR1-3 and serine-glycine rich junction (SRJ). All other mutations were located in SRJ except p.P140S.</p

Position of variations

<p>Position of variations</p