Springer Science and Business Media LLC

Kefir, a traditional fermented milk product, is considered as a rich source of biofunctional compounds like peptides that are beneficial to human health. The objective of this study was to extract water-soluble peptides from cow and ewe milk kefir (produced by local kefir grains of Semnan, Iran) and investigate their antioxidant and antimicrobial activities. Evaluation of the chemical composition of the cow and ewe pasteurized milk and kefir showed a decrease in protein, total carbohydrate, and lactose content. There was no difference between the mineral profile of fresh milk and kefir. The count of lactic acid bacteria was almost stable during the 28 days of storage period. From the 1st to the 28th day of storage, the population of Lactococci (LC) was 11.5–12.1 and 11.4–12.2 log10 CFU/mL for cow and ewe milk kefir, respectively. A great antioxidant potential (DPPH and ABTS assay) was observed for the peptides extracted from cow and ewe kefir (at a concentration of 1.25 mg/mL) on day 28 storage. Also, ewe kefir peptides had higher antioxidant activity than peptides of cow kefir. The antimicrobial activity of water-soluble peptides of ewe milk kefir was superior to those of cow milk kefir (at the same concentration of 20 mg/mL). The results of this work expressed great functional properties of peptides derived from ewe milk produced by local kefir grains of Semnan.

Hepatitis C virus (HCV) infection is a serious global health problem and a cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Bioinformatics software has been an effective tool to study the HCV genome as well as core domains. Our research was based on employing several bioinformatics software applications to find important mutations in domain 1 of core protein in Iranian HCV infected samples from 2006 to 2017, and an investigation of general properties, B-cell and T-cell epitopes, modification sites, and structure of domain 1. Domain 1 sequences of 188 HCV samples isolated from 2006 to 2017, Iran, were retrieved from NCBI gene bank. Using several tools, all sequences were analyzed for determination of mutations, physicochemical analysis, B-cell epitopes prediction, T-cell and CTL epitopes prediction, post modification, secondary and tertiary structure prediction. Our analysis determined several mutations in some special positions (70, 90, 91, and 110) that are associated with HCC and hepatocarcinogenesis, efficacy of triple therapy and sustained virological response, and interaction between core and CCR6. Several B-cell, T-cell, and CTL epitopes were recognized. Secondary and tertiary structures were mapped fordomain1 and core proteins. Our study, as a first report, offered inclusive data about frequent mutation in HCV-core gene domain 1 in Iranian sequences that can provide helpful analysis on structure and function of domain 1 of the core gene.

Currently bioactive peptides are the main focus in attempts to identify novel angiotensin-converting enzyme inhibitors (ACEIs) for the treatment of hypertension due to their fewer side effects. In the present study we aimed to investigate the antihypertensive activity of 4 synthetic tripeptides Ornithyl-Hydroxyprolyl-Proline (Orn-Hyp-Pro OhPP) Ornithyl-Prolyl-Hydroxyproline (Orn-Pro-Hyp OPhP) Citrullyl-Hydroxyprolyl-Proline (Cit-Hyp-Pro ChPP) and Citrullyl-Prolyl-Hydroxyproline (Cit-Pro-Hyp CPhP) in vitro and in vivo. The ACE inhibitory activity and mode were analyzed with a modified spectrophotometric method and Lineweaver-Burk plots respectively. It showed that peptide Citn-Hyp-Pro (ChPP) exhibited the highest inhibition potency with an IC50 value of 40.48 μM and displayed a competitive inhibition of ACE. Molecular docking simulations suggested that ChPP could form several critical hydrogen bonds with the major residues His353 and His513 located in the S2 active site of ACE. The systolic blood pressure of spontaneously hypertensive rats monitored by the tail-cuff method was significantly reduced by 15.54% at 4 h after oral administration of 20 mg/kg BW ChPP. Also ChPP remarkably downregulated the angiotensin II receptor type 1 (agtr1) and miR-132/-212 levels in SHRs which was similar to that observed in SHRs treated with captopril. It showed us that the tripeptide ChPP could be explored as a promising ACE inhibitor (ACEI) for treatment of hypertension.

Drug resistance by microbes is becoming a major concern in the field of medicine. The oligo-dynamic nature of silver nanoparticles (AgNPs) is capable of killing even the multi-drug resistant organisms. In the current study, AgNPs were synthesized from cell free supernatant of Lactic Acid Bacteria (LAB), isolated from the gut of marine fishes viz., Lutjanus campechanus, Dendrobranchiata and Gerres subfasciatus. The synthesis of AgNPs was evaluated by the change in colour from pale-yellow to dark brown. Further, characterization based on Scanning Electron Microscopy and particle size analysis revealed the structure alignment, shape and average size of the synthesized AgNPs. X-Ray Diffraction revealed the unique diffraction peaks of the synthesized AgNPs. Fourier Transform Infra-Red spectroscopy confirmed the presence of carboxyl, amine and hydroxyl groups. The activity of the synthesized AgNPs was evaluated against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. Results revealed a substantial increase in activity against the tested bacteria. Further, successful binding of AgNPs with collagen asserts its application as a potential material with biological utility.

Fish scales-derived collagen peptides (CPs) are characterized by their specific amino acid composition with a high concentration of glycine, proline and hydroxyproline. These amino acids have been known to exert beneficial effects on human skin. The aim of the present study was to examine the effects of collagen peptides obtained from fish scales on changes in periorbital wrinkles, facial skin hydration, and skin elasticity in healthy women aged 30–60 years. In the present randomized, placebo-controlled, double-blind trial, 71 subjects consumed a 20 mL beverage containing 3000 mg of CPs or placebo once per day over 12 weeks. Significant decreases in periorbital wrinkles (p < 0.05) were observed in the treatment group after 12 weeks of CPs ingestion compared to the control group. This study also demonstrated a consistent trend of enhanced facial skin moisture (p < 0.001) and skin elasticity (p < 0.001) by dietary intake of CPs without any side effects or adverse events. These findings indicate that fish-derived CPs hold great promise as a natural supplement with cutaneous anti-aging properties.

The Nuclear Mitotic Apparatus protein (NuMA) is a nuclear protein that plays critical role in the mitosis as an organizer of the mitotic spindles and is a prevalent protein in malignant urothelial cells which makes it an appropriate marker for bladder cancer early detection. In this study, NuMA protein sequence was obtained from Genbank based on literature studies. Immunodominant epitopes which are located between residues 1 and 300 were reverse translated and optimized for expression in E. coli cells. Final sequence was then synthesized, cloned into pGS21a vector and expressed in BL21 cells. The expressed protein was examined by SDS-PAGE and also Western blotting to confirm expression by using anti-His antibody-HRP. The protein was then purified using nickel chromatography column and 18 mg of purified truncated NuMA was obtained from 1 l of bacterial culture. ELISA was performed to evaluate the reactivity of commercial anti-NuMA antibody towards truncated protein. Finally, the antigen was injected and polyclonal antibody was produced in rabbits. The immunoglobulins of antiserum were precipitated by ammonium sulfate and purified by DEAE–Cellulose chromatography column. To evaluate antibody accuracy, it was tested on HEp-2 cells with permeable nucleus membranes fixed on slides. The antibody detected NuMA protein in the nuclei of HEp-2 cells, which means it can have diagnostic value in the bladder cancer patients. The truncated NuMA is a suitable protein as a single multiepitope antigen that further may be used for studies to develop assays for early diagnosis of bladder cancer.

In this minireview paper it has been elucidated that the proposal of pseudo amino acid components represents a very important milestone for the disciplines of proteome and genome. This has been concluded by observing and analyzing the developments in the following six different sub-disciplines: (1) proteome analysis; (2) genome analysis; (3) protein structural classification; (4) protein subcellular location prediction; (5) post-translational modification (PTM) site prediction; (6) stimulating the birth of the renowned and very powerful 5-steps rule.

A well-organized protocol has been developed for high frequency root germination from the seed of Canavalia ensiformis on Murashige and Skoog (MS) medium. Surprisingly, the seeds that were grown on the MS medium having no growth hormone showed the best response. Roots of 30 days old aseptic seedling were homogenized and a lectin from them was purified on Sephadex G-50 affinity column. The finding that final product is a pure lectin was confirmed by specific hemagglutinating property. The final root lectin yield was 0.6% and eluted as a single peak. Root lectin specific activity was 50 times more than the seed lectin. Sugar specificity activity by hemagglutination-inhibition assay indicated that lectin belongs to glucose/mannose-specific group. Interestingly, the lectin was found to be 25 kDa, similar to molecular mass of Concanavalin A purified from seed of C. ensiformis, as revealed by SDS–PAGE. Thus, Concanavalin A from either source can be used for development of transgenic crops that are capable of expressing lectin gene and hence can efficiently perform biological nitrogen fixation by giving rise to nodules in their root. The advantage of this method is that purification of Concanavalin A in tissue culture conditions is easier, handy and is less time consuming.

Alterations in thyroid function are associated with changes in body weight, metabolism, and low-grade inflammation abnormal thyroid function may be associated with disturbances in the production of adipokines also. Although there have been studies showing changes in visfatin levels in thyroid dysfunction, exact relationship between them was still unclear. Our aim was to evaluate serum concentrations of visfatin in patients with subclinical thyroid dysfunction before and after normalization of thyroid function tests. The study included 43 patients (mean age 50.1 ± 10.6 years) with subclinical hypothyroidism. Serum insulin, visfatin, TSH, free T4 (FT4) and free T3 (FT3) levels of subjects were analyzed. Visfatin levels were measured in all patients before starting therapy and after normalization of thyroid function. Serum visfatin levels of subclinical hypothyroid patients were 0.99 ± 0.45 and they were similar after normalization of thyroid function (p = 0.394). Serum visfatin levels were negatively correlated with FT4 levels before treatment (r = −0.329 p < 0.05). There was no significant correlation between serum levels of visfatin and the serum levels of TSH and FT3. Serum visfatin levels did not correlate with insulin, fasting blood glucose, total cholesterol, HDL cholesterol, LDL cholesterol and triglyceride levels. In this study, it was shown visfatin levels did not change after replacement therapy in patients with subclinical hypothyroidism. Subclinical hypothyroid state may be an earlier stage regarding the changes of adipocytokines specifically the visfatin secretion as seen in overt hypothyroidism.

Since its discovery in 1989, there has been extensive research on endothelin (ET)-1 physiology, as well as pathology. Accordingly, there is considerable research on the discovery of therapeutics based around ET-1, amongst which current treatment options include endothelin receptor antagonists. These target the ET-1 receptors, which are G-protein–coupled receptors (GPCRs). We have effectively developed a soluble form of a GPCR that binds to ligands, by constructing a fusion polypeptide of different endothelin receptor ligand binding domains. Phage experiments identified strong binders to ET-1. We then constructed Fc-fusions of the top binders and further binding assays revealed a KD of 21.2 nM for the Fc-ETtr1 construct and KD of 77.3 nM for the Fc-ETtr2 construct. These constructs are soluble and have the ability to bind and potentially sequester elevated ET-1 levels that are prevalent in different diseases. These results provide a novel approach to targeting GPCR–binding ligands, and thereby to contribute to a very important class of therapeutics.