Expression of Penaeidins 3a in Transgenic Duckweed Confers Antibacterial Ability

Background With the development of aquaculture, fish and shrimp diseases have been paid more and more attention in the world. How to improve the immunity of aquatic animals was an urgent problem to be solved. Duckweed (Lemnacecae), as a eukaryote, could be an ideal feedstock for the production of antimicrobial peptides. Result Penaeidins 3a (Pen 3a) from Litopenaeus vannamei was expressed under the control of CaMV-35S promoter in duckweed, Lemna turionifera 5511. Bacteriostatic test by Pen3a duckweed extract showed the antibacterial activity against Escherichia coli and Staphylococcus aureus. Transcriptome analysis of WT and Pen3a duckweed showed different results, and the protein metabolic process was the most up-regulated DEGs. In Pen 3a transgenic duckweed, the expression of sphingolipid metabolism and phagocytosis process-related genes have been significantly up-regulated. Quantitative proteomics suggested a remarkable difference in protein enrichment in metabolic pathways. Conclusion Our study provide a novel solution on aquaculture and water purification. The Pen 3a transgenic duckweed extraction inhibit the growth of gram-negative bacteria, gram-positive bacteria, which could be applied to control the bacteria in lake. The results could lay the foundation for the subsequent production of antibiotics.

Cycles of transcription and local translation consolidate Arc protein at dendritic sites

Maintenance of long-term memory requires transcription and translation of activity-regulated genes. Many of these are immediate early genes (IEGs) with short-lived mRNAs and proteins, often decaying rapidly after stimulation. It remains unknown how an IEG with rapid mRNA and protein turnover can impact long-lasting changes at the synapses. Here, we imaged the long-term transcription and translation dynamics of an IEG, Arc, important for memory consolidation, after stimulation in individual neurons. We demonstrated that the gene underwent reactivation in the same neuron without an additional stimulus, often at the same allele. The cycling of Arc transcription required protein synthesis. Indeed, phases of Arc translation were coordinated with the transcription cycles. Arc mRNAs from later cycles preferentially localized at dendritic sites marked by previous Arc protein, thereby creating hubs of local protein enrichment, potentially maintained by cycles of dendritic translation. These results identify novel characteristics of an IEG and provide insights into how unstable proteins may be sustained over the long-time scale of memory consolidation.

A Simple and Low-Cost Method for Fabrication of Polydimethylsiloxane Microfludic Chips

The conventional fabrication methods for enrichment microfluidic devices require cleanroom, which are costly and time-consuming. Developing a facile and low-cost method to fabricate microfluidic chips could stimulate the progress of the applications of those chips. Here, we present an easy method for fabrication of a complete PDMS (Polydimethylsiloxane) microfluidic chip used for ion and protein enrichment. The method consists of three main fabrication steps: PDMS microchannels ablation by co2 laser, nation membrane deposition, and oxygen plasma assist bonding under pressure. To fabricate a desired microchannel, the laser ablation parameters, containing laser power and ablation speed, were analyzed. The parameters for oxygen plasma assist bonding were also investigated to improve the bonding quality of the chips (low dimension loss and high bonding strength). The following Rhodamine B enrichment tests demonstrate that the presented method allows fabrication of microfluidic chips with precise dimensions and leakage free.

Dia- and Rok-dependent enrichment of capping proteins in a cortical region

Rho signaling with its major targets the formin Dia, Rho kinase (Rok) and non-muscle myosin II control turnover, amount and contractility of actomyosin. Much less investigated has been a potential function for the distribution of F-actin plus and minus ends. In syncytial Drosophila embryos Rho1 signaling is high between actin caps, i. e. the cortical intercap region. Capping protein binds to free plus ends of F-actin to prevent elongation of the filament. Capping protein has served as a marker to visualize the distribution of F-actin plus ends in cells and in vitro. Here, we probed the distribution of plus ends with capping protein in syncytial Drosophila embryos. We found that Capping proteins are specifically enriched in the intercap region similar to Dia and MyoII but distinct from overall F-actin. The intercap enrichment of Capping protein was impaired in dia mutants and embryos, in which Rok and MyoII activation was inhibited. Our observations reveal that Dia and Rok/MyoII control Capping protein enrichment and support a model that Dia and Rok/MyoII control the organization of cortical actin cytoskeleton downstream of Rho1 signaling.

The polarity protein Par3 coordinates positively self-renewal and negatively invasiveness in glioblastoma

Glioblastoma (GBM) is a brain malignancy characterized by invasiveness to the surrounding brain tissue and by stem-like cells, which propagate the tumor and may also regulate invasiveness. During brain development, polarity proteins, such as Par3, regulate asymmetric cell division of neuro-glial progenitors and neurite motility. We, therefore, studied the role of the Par3 protein (encoded by PARD3) in GBM. GBM patient transcriptomic data and patient-derived culture analysis indicated diverse levels of expression of PARD3 across and independent from subtypes. Multiplex immunolocalization in GBM tumors identified Par3 protein enrichment in SOX2-, CD133-, and NESTIN-positive (stem-like) cells. Analysis of GBM cultures of the three subtypes (proneural, classical, mesenchymal), revealed decreased gliomasphere forming capacity and enhanced invasiveness upon silencing Par3. GBM cultures with suppressed Par3 showed low expression of stemness (SOX2 and NESTIN) but higher expression of differentiation (GFAP) genes. Moreover, Par3 silencing reduced the expression of a set of genes encoding mitochondrial enzymes that generate ATP. Accordingly, silencing Par3 reduced ATP production and concomitantly increased reactive oxygen species. The latter was required for the enhanced migration observed upon silencing of Par3 as anti-oxidants blocked the enhanced migration. These findings support the notion that Par3 exerts homeostatic redox control, which could limit the tumor cell-derived pool of oxygen radicals, and thereby the tumorigenicity of GBM.

Protein Enrichment of Donor Breast Milk and Impact on Growth in Very Low Birth Weight Infants

Protein content is often inadequate in donor breast milk (DBM), resulting in poor growth. The use of protein-enriched target-pooled DBM (DBM+) has not been examined. We compared three cohorts of very low birth weight (VLBW) infants, born ≤ 1500 g: DBM cohort receiving > 1-week target-pooled DBM (20 kcal/oz), MBM cohort receiving ≤ 1-week DBM, and DBM+ cohort receiving > 1-week DBM+. Infants followed a standardized feeding regimen with additional fortification per clinical discretion. Growth velocities and z-scores were calculated for the first 4 weeks (n = 69 for DBM, 71 for MBM, 70 for DBM+) and at 36 weeks post-menstrual age (n = 58, 64, 59, respectively). In total, 60.8% MBM infants received fortification >24 kcal/oz in the first 30 days vs. 78.3% DBM and 77.1% DBM+. Adjusting for SGA, length velocity was greater with DBM+ than DBM in week 1. Average weight velocity and z-score change were improved with MBM compared to DBM and DBM+, but length z-score decreased similarly across all groups. Incidences of NEC and feeding intolerance were unchanged between eras. Thus, baseline protein enrichment appears safe in stable VLBW infants. Weight gain is greatest with MBM. Linear growth comparable to MBM is achievable with DBM+, though the overall length trajectory remains suboptimal.