Protein and Peptide Letters - Volume 26, Issue 4, 2019

Seminal plasma proteins contributed by secretions of accessory glands plays a copious role in fertilization. Their role is overlooked for decades and even now, as Artificial Reproduction Techniques (ART) excludes the plasma components in the procedures. Recent evidences suggest the importance of these proteins starting from imparting fertility status to men, fertilization and till successful implantation of the conceptus in the female uterus. Seminal plasma is rich in diverse proteins, but a major part of the seminal plasma is constituted by very lesser number of proteins. This makes isolation and further research on non abundant protein a tough task. With the advent of much advanced proteomic techniques and bio informatics tools, studying the protein component of seminal plasma has become easy and promising. This review is focused on the role of seminal plasma proteins on various walks of fertilization process and thus, the possible exploitation of seminal plasma proteins for understanding the etiology of male related infertility issues. In addition, a compilation of seminal plasma proteins and their functions has been done.

Background: Molecular imaging of cancer cells using effective drug targeting systems are most interested research area in recent years. Albumin protein is a soluble and most abundant protein in circulatory system. It has a ligand-binding function and acts as a transport protein. Researchers are interested in developing albumin based nanostructured specific anti-tumor drugs in cancer therapy. Pancreatic cancer treatment or drug design for targeted pancreatic cancer cell has great importance due to it has a high mortality rate comparing other cancer types. Objective: In this article, our goal is to develop new targeting nanoparticles based on the conjugation of albumin and Hyaluronic Acid (HA) for pancreatic cancer cells. Method: In this article, we proposed a new technique for conjugation of albumin (BSA) and HA in nano formation. Firstly, cationic BSA is synthesized. Then, BSA-HA conjugation is obtained by interacted cationic BSA with 1000 ppm HA. Secondly, nano BSA-HA particles and nano BSA particles were synthesized according to AmiNoAcid Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method which provides a special cross-linking strategy for biomolecules using ruthenium-based amino acid monomer haptens. After characterization studies, in vitro cytotoxic activity of synthesized nano BSA-HA particles were determined for PANC-1 ATCC® CRL146 cells. Results: According to the data, nano BSA and nano BSA-HA particles synthesized uniquely using special ruthenium-based amino acid decorated cross-linking agent, (MATyr)2-Ru-(MATyr)2.based on ANDOLUCA method. Characterization results showed that there was not any change in protein folding structures during nano formation process. In addition, nano protein particles gained fluorescence feature. When interacting synthesized nano BSA and nano BSA-HA particles with pancreatic cells, it was found that BSA nanoparticles were usually around cells and membranes, but BSA-HA nanoparticles were identified around the cells, in the cytoplasm inside the cell, and next to the cell nucleus. So, nano BSA-HA particles could be used as cancer cell imaging agent for PANC-1 ATCC® CRL146 cells. Conclusion: The satisfactory conclusion of this study is that synthesized nano BSA-HA particles are fundamental materials for targeting pancreatic cancer cells due to HA receptors located on pancreatic cancer cells and imaging agents due to fluorescence feature of the BSA-HA nanoparticles.

Introduction: Several studies have been conducted to find new biomarkers for the discrimination of Latent Tuberculosis Infection (LTBI) from active TB (ATB); however, their findings are inconsistent. The aim of the current study was to evaluate the potential of in vitro antigenspecific expression of Monocyte Chemotactic Protein (MCP)-2 for discrimination of ATB and LTBI after stimulation of whole blood with PE35 and PPE68 recombinant proteins. Materials and Methods: The recombinant PE35 and PPE68 proteins were evaluated at a final concentration of 5 μg/ml by a 3-day whole blood assay. Secreted MCP-2 from the culture supernatants were measured by commercially available Human MCP2 ELISA Kit. The diagnostic performance of MCP-2 was ascertained by Receiver Operator Characteristic (ROC) curve and measuring the Area Under the Curve (AUC) and their 95% confidence intervals (CI). Cut-offs was estimated at various sensitivities and specificities and at the maximum Youden’s index (YI), i.e. sensitivity specificity–1. Results: The median MCP-2 response to both PE35 and PPE68 in those with LTBI was significantly higher than patients with ATB. The discrimination performance of MCP-2 response following stimulation of PE35 (assessed by AUC) between LTBI and patients with ATB was 0.98 (95%CI: 0.94-1.00). Maximum discrimination was reached at a cut-off of 86pg/mL with 100% sensitivity and 97% specificity. The highest sensitivity and specificity was obtained using cut off 58 pg/mL following stimulation with PPE68 (100% and 90%, respectively; AUC: 0.94, 95%CI: 0.85- 1.00). Conclusion: MCP-2 induced by PE35 and PPE68 shows good discriminatory power for discrimination of ATB and LTBI. Additional studies with a larger sample size are needed to confirm the advantage of this marker, alone or combined with other markers; however, these findings present a promising method, which can discriminate between ATB and LTBI.

Background: There are few studies on the autophagy and inflammatory effects of soy peptides on the inflammatory cell model. Further insight into the underlying relationship of soybean peptides and autophagy needs to be addressed. Therefore, it is worthwhile investigating the possible mechanisms of soybean peptides, especially autophagy and the inflammatory effects. Objective: In this study, we used a RAW264.7 cell inflammation model to study the inhibitory effect and mechanism of soybean peptide QRPR on inflammation. Methods: We used LPS-induced inflammation model in RAW264.7 cells to study the inhibitory effect and mechanism of soybean peptide QRPR on inflammation. First, Cell viability was determined by cell activity assay. Subsequently, the concentrations of the inflammatory cytokines IL-6 and TNF-α were measured by ELISA. IL-6, TNF-α, Beclin1, LC3, P62, PIK3, AKT, p-AKT, pmTOR and mTOR protein expression were detected by western-blot. PIK3, AKT and mTOR gene expression level were quantified by quantitative real-time PCR. Double-membrane structures of autophagosomes and autolysosomes were observed by transmission electron microscopy. The PI3K/AKT/mTOR signaling pathway in LPS-induced RAW264.7 cells was speculated when the autophagy was activated. Results: The results showed that QRPR activates autophagy in the inflammatory cell model and that the inhibitory effect of QRPR on inflammation is reduced after autophagy was inhibited. Western- blot and real-time PCR results indicated that QRPR activates autophagy in LPS-induced RAW264.7 cells by modulating the PI3K/AKT/mTOR signaling pathway, and it shows a significant time dependence. Conclusion: This study indicated that the soybean peptide QRPR activates autophagy and attenuates the inflammatory response in the LPS-induced RAW264.7 cell model.