Phenylalanine Rich Diet (Vicia faba L.) Enhances the Expression of Dopamine Receptor D3 (DRD3) Gene in Rabbits

Dopamine is a neurotransmitter hormone for pleasure and reward. It is synthesized only in the brain cells of human and animals and responsible for the regulation of behavior, mood, memory, cognitive, flexible movement, the body weight, and other important functions. The Dopamine Receptor D3 (DRD3) is the most important receptor for dopamine. In this investigation, expression of DRD3 gene was studied in rabbits fed on supplemented diet of dry and fresh faba bean (Vicia faba L. Sakha 3). DRD3 gene (≈ 1200 bp) in control and treated rabbits were PCR amplified, sequenced and aligned with reference gene (Acc. No XM_017346708.1). High genetic similarity values were detected among all sequences. DRD3 gene sequences of control, fresh and dry faba bean fed rabbits were deposited in the GenBank with accession numbers MZ714134, MZ714135 and MZ714136 respectively. Direct estimation of blood phenylalanine (Phe) amino acid indicated that feeding rabbits on dry faba bean reflected the highest level of Phe in the rabbit’s blood. Quantitative RT-qPCR analysis showed that DRD3 gene was over expressed after feeding rabbits on dry faba bean form compared with feeding on green form and control. Thus, diet rich with phenylalanine like Sakha3 (dry and fresh forms) enhance gene expression of DRD3 gene. However, diet doesn't affect the DRD3 gene sequence and structure. In a conclusion, our findings indicated a direct effect of faba bean supplemented diet on increasing DRD3 expression levels which improve the life quality for human.

What’s Wrong in a Jump? Prediction and Validation of Splice Site Variants

Alternative splicing (AS) is a crucial process to enhance gene expression driving organism development. Interestingly, more than 95% of human genes undergo AS, producing multiple protein isoforms from the same transcript. Any alteration (e.g., nucleotide substitutions, insertions, and deletions) involving consensus splicing regulatory sequences in a specific gene may result in the production of aberrant and not properly working proteins. In this review, we introduce the key steps of splicing mechanism and describe all different types of genomic variants affecting this process (splicing variants in acceptor/donor sites or branch point or polypyrimidine tract, exonic, and deep intronic changes). Then, we provide an updated approach to improve splice variants detection. First, we review the main computational tools, including the recent Machine Learning-based algorithms, for the prediction of splice site variants, in order to characterize how a genomic variant interferes with splicing process. Next, we report the experimental methods to validate the predictive analyses are defined, distinguishing between methods testing RNA (transcriptomics analysis) or proteins (proteomics experiments). For both prediction and validation steps, benefits and weaknesses of each tool/procedure are accurately reported, as well as suggestions on which approaches are more suitable in diagnostic rather than in clinical research.

An Information Gain-based Method for Evaluating the Classification Power of Features Towards Identifying Enhancers

Background: Enhancers are cis-regulatory elements that enhance gene expression on DNA sequences. Since most of enhancers are located far from transcription start sites, it is difficult to identify them. As other regulatory elements, the regions around enhancers contain a variety of features, which can help in enhancer recognition. Objective: The classification power of features differs significantly, the performances of existing methods that use one or a few features for identifying enhancer vary greatly. Therefore, evaluating the classification power of each feature can improve the predictive performance of enhancers. Methods: We present an evaluation method based on Information Gain (IG) that captures the entropy change of enhancer recognition according to features. To validate the performance of our method, experiments using the Single Feature Prediction Accuracy (SFPA) were conducted on each feature. Results: The average IG values of the sequence feature, transcriptional feature and epigenetic feature are 0.068, 0.213, and 0.299, respectively. Through SFPA, the average AUC values of the sequence feature, transcriptional feature and epigenetic feature are 0.534, 0.605, and 0.647, respectively. The verification results are consistent with our evaluation results. Conclusion: This IG-based method can effectively evaluate the classification power of features for identifying enhancers. Compared with sequence features, epigenetic features are more effective for recognizing enhancers.

Electrospun poly(l-lactide-co-caprolactone)–collagen–chitosan vascular graft in a canine femoral artery model

(P(LLA-CL)–COL–CS) composite vascular grafts could effectively improve patency rate, promote tissue regeneration, and enhance gene expression.

Diglycidyl Esters Cross-Linked with Low Molecular Weight Polyethyleneimine for Magnetofection

Magnetic polyethyleneimine (PEI) complexes have demonstrated to be simple and efficient vectors for enhancing gene transfection. However, the high cytotoxicity of PEI restricts its further application in vivo. In this study, we synthesized several low cytotoxicity biodegradable cationic polymers derived from PEI (Mw 600) linked with diglycidyl tartrate (DT-PEI) or its analogues (diglycidyl succinate (DS-PEI) and diglycidyl malate (DM-PEI); D-PEIs for all 3 polymers). Moreover, a type of biocompatible magnetic nanoparticles (MNPs) with negative charges was prepared to assemble with D-PEIs/DNA complexes via electrostatic interactions. The magnetic ternary complexes have appropriate sizes of 120–150 nm and zeta potential values of ~20–25 mV. The transfection ability and cell viability of D-PEIs increased as the amount of hydroxyl groups increased in the repeat unit, which indicated that increasing the hydroxyl number in the backbone of D-PEIs can enhance gene expression and decrease cytotoxicity in A549 cells. Magnetofection of DT-PEI showed similar transfection efficiency with 30 min incubation; in contrast, the standard incubation time was 4 h. All three magnetic complexes displayed lower cytotoxicity when compared with those of PEI complexes in COS-7 and A549. These results indicated that these series of magnetic PEI derivatives complexes could be potential nanocarriers for gene delivery.