Protein hydrolysates produced from different food sources exhibit therapeutic potential and can be used in the management of chronic diseases. This study was targeted to optimise the conditions for the hydrolysis of stone fish protein to produce antioxidant hydrolysates using central composite design (CCD) by response surface methodology (RSM). The stone fish protein was hydrolysed under the optimum predicted conditions defined by pH (6.5), temperature (54°C), E/S ratio (1.5%), and hydrolysis time (360 min). The hydrolysates were then evaluated for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) scavenging activity and ferrous ion- (Fe2+-) chelating activity. Results validation showed no significant difference between the experimental values of DPPH• scavenging activity (48.94%) and Fe2+-chelating activity (25.12%) obtained at 54.62% degree of hydrolysis (DH) compared to their corresponding predicted values of 49.79% and 24.08% at 53.08% DH, respectively. The hydrolysates demonstrated non-Newtonian behavior (n < 1) with stronger shear-thinning effect and higher viscosities at increasing concentration. Thus, RSM can be considered as a promising strategy to optimise the production of stone fish protein hydrolysates containing antioxidant peptides. It is hoped that this finding will enhance the potential of stone fish protein hydrolysates (SHs) as therapeutic bioactive ingredient in functional foods development.
In this work, we describe results of the reproductive health monitoring studies in Mytilus galloprovincialis following spermatozoa hsp70 expression and protamine-like protein properties. Mussels control (ctr) were released within cages for 30 days in three different marine sites near Naples (Campania, Italy): Bagnoli south (BAs) and Bagnoli north (BAn), both close to a disposal metallurgical factory and in Capo Miseno (CM). Studies of hsp70 gene expression carried out, by RT-qPCR, in mussel spermatozoa have shown varied expression levels, particularly 5, 13, and 15-fold more than ctr in CM, BAs, and BAn, respectively, indicating highest involvement of stress proteins in spermatozoa of mussels in Bagnoli. In order to evaluate the possible risk on Mytilus galloprovincialis sustainability loss, electrophoretic analyses were performed on protamine-like proteins (PL) of collected spermatozoa. The results showed that CM PL were apparently unaltered with respect to ctr PL, while BAs and BAn PL appeared in part in the form of peptides and in part as bands with low mobility. Further, CM and BAs PL showed, by electrophoretic mobility shift assay, a decrease in DNA binding ability and a change in their DNA binding mode. The results of this investigation show the usefulness of the study of alterations of spermatozoa hsp70 expression and protamine-like protein properties for eco-toxicological evaluation using Mytilus galloprovincialis as a bioindicator.
Winged bean seed (WBS) is an underutilized tropical crop. The current study evaluates its potential to reduce blood pressure (BP) in spontaneously hypertensive rats and finds that it reduces BP significantly, in a dose-dependent manner. Five peptides with the sequences, RGVFPCLK, TQLDLPTQ, EPALVP, MRSVVT and DMKP, have been characterized in terms of their stability against ACE via in vitro and in silico modelling. All peptides exhibited IC50 values between 0.019 and 6.885 mM and various inhibitory modes, including substrate, prodrug and true inhibitor modes. The toxicity status of non-Current Good Manufacturing Practice (non-CGMP) peptides is evaluated and the results show that such peptides are toxic, and thus are not suitable to be tested in animals, particularly in repeated-dose studies. In short, WBS hydrolysate demonstrated in vitro ACE inhibitory properties and in vivo blood pressure lowering efficacy in rat models, fostering its potential as a functional food ingredient. Non-CGMP grade peptides are toxic and unfit for testing in animal models.
Serratia marcescens subsp.sakuensisstrain K27 was isolated from sponge (Haliclona amboinensis). The genome of this strain consists of 5,325,727 bp, with 5,140 open reading frames (ORFs), 3 rRNAs, and 67 tRNAs. It contains genes for the production of amylases, lipases, and proteases. Gene clusters for the biosynthesis of nonribosomal peptides and thiopeptide were also identified.
Cancer still presents enormous challenges in the medical world. Currently, the search for
anticancer compounds has garnered a lot of interest, especially in finding them from the natural
sources. In this study, by using Sulforhodamine B (SRB) colorimetric assay, compounds,
extracted from supermeal worm (Zophobas morio) larvae using two types of acidified organic
solvent (ethanol and isopropanol), were shown to inhibit the growth of a breast cancer line,
MCF-7. A comparative study of the effect was carried out on a normal cell line, Vero. Results
showed that, the two types of extracts inhibits growth of MCF-7 cell at varying degrees, on
the other hand, have much less effect on Vero cell. Extracts analysed by UV-vis spectroscopy,
showed peaks in the range of 260 to 280 nm, inferring the presence of aromatic amino acids,
whereas the highest peak of 3.608 AU at 230 nm indicates the presence of peptide bonds. By
Raman spectroscopy, peaks are observed at 1349 cm-1, 944 cm-1 and 841 cm-1 indicating the
presence of Tyr, Try and Gly, confirming the UV-vis analyses. All results of analyses implied
that the anticancer compounds contain peptides.
Haemoglobin E (Hb E) is a variant of structurally abnormal haemoglobin that can be found very commonly in the Asian countries particularly the Southeast Asian [1]. [H1] Alpha thalassaemia is a red cell disorder which is caused by deletion or mutation of one or more of the four alpha globin genes leading to absence or decrease in production of alpha globin peptides [2]. This disorder is far more common in South East Asian regions and in Malaysia itself, and the gene frequency is about 4.1% [2]. The interactions of Hb E and alpha thalassaemia are evident in Kelantan which is bordered by southern Thailand. Using capillary electrophoresis (CE), a reduction of Hb E level is noticed as compared to Hb E heterozygotes. DNA analysis should be done to determine the presence of concurrent alpha thalassaemia variant. This study was done to evaluate haematological parameters using automated blood counters, morphology of red cells, Hb separation and quantitation of Hb fractions using CE and molecular analysis for alpha thalassemia. The study also aimed to discover cut off point of Hb E level in heterozygous Hb E patients with concurrent deletional alpha thalassaemia by CE.
Alzheimer's disease (AD) is the most common form of dementia and has affected millions of individuals worldwide. The hallmarks of AD include the amyloid beta plaque deposits, tau neurofibrillary tangles, altered neuronal signaling, alongside decline in memory and cognitive functions. Conventional drug therapies do exist, such as donepezil or aducanumab, but these drugs mostly focus on halting AD progression instead of causing a reversal within the disease. In an effort to ameliorate and ultimately cure AD, researchers have delved into viral-mediated gene therapy to fix this disease from its root molecular causes. To date, adeno-associated virus and lentiviral vectors have remained the most vastly studied among other viral vectors to combat AD. These vectors could be employed alongside various genetic materials based on the types of processes we want to alter to yield a positive effect, such as disruption of amyloidogenic pathway, neuroprotection and lipid metabolism pathways. Recent studies and trials were reviewed in this article, highlighting their clinical significance, differences and limitations between each method. By learning from the different combinations and possibilities of viral-mediated gene transfer, researchers would then get a step closer in ameliorating symptoms and possibly in curing AD.
Protease is an enzyme that catalysed the hydrolysis of protein into peptide. Application of protease in industry has been linked with cost effective substrates and complex of enzyme-substrate stability. Molecular docking approach has identified casein as a preference substrates. However, lack of data on casein mode of binding to protease and enzyme stability represents a limitation for its production and structural optimization. In this study, we have used a molecular dynamic (MD) to examine the stability of complex enzyme-substrate of protease from Bacillus lehensis G1. The 3D structure of protease (BleG1_1979) was docked with substrate casein using AutoDock Vina. Structural analysis of the substrate-binding cleft revealed a binding site of casein was predominantly at the hydrophobic region of BleG1_1979. The MD of complex BleG1_1979-casein was tested with two temperatures; 298 K and 310 K using GROMACS v5.1.4. MD simulation showed a stable behaviour of BleG1_1979 over the 20 ns simulation period. The molecular docking and MD simulation suggested that the production of protease from B. lehensis G1 by utilization of casein and the stability of complex protease-casein could be a potential application to generate a cost effective enzyme to be develop for industrial use.
Actinopyga lecanora, as a rich protein source was hydrolysed to generate antibacterial bioactive peptides using different proteolytic enzymes. Bromelain hydrolysate, after 1 h hydrolysis, exhibited the highestantibacterial activities against Pseudomonas aeruginosa, Pseudomonas sp., Escherichia coli and Staphylococcus aureus. Two dimensional fractionation strategies, using a semi-preparative RP-HPLC and an isoelectric-focusing electrophoresis, were applied for peptide profiling. Furthermore, UPLC-QTOF-MS was used for peptides identification; 12 peptide sequences were successfully identified. The antibacterial activity of purified peptides from A. lecanora on P. aeruginosa, Pseudomonas sp., E. coli and S. aureus was investigated. These identified peptides exhibited growth inhibition against P. aeruginosa, Pseudomonas sp., E. coli and S. aureus with values ranging from 18.80 to 75.30%. These results revealed that the A. lecanora would be used as an economical protein source for the production of high value antibacterial bioactive peptides.
The emergence of bacterial resistance against conventional antibiotics and the growing interest in developing alternative, natural antibacterial agents have prompted the search for plant-derived antibacterial peptides in recent decades. Different classes of endogenous antibacterial peptides have been identified from various plant species. Moreover, protein hydrolysates and hydrolysate-derived peptides with potent antibacterial effects have also been identified from numerous plant sources. Antibacterial peptides are often cationic and amphipathic, consisting of fewer than 100 amino acids. They are able to disrupt bacterial membrane integrity via pore formation and/or compromise bacterial metabolic processes. In this review, we summarize current knowledge on the characteristics and modes of action of antibacterial peptides, as well as salient points concerning the production of antibacterial protein hydrolysates from plant proteins. Examples of plant-derived antibacterial hydrolysates and peptides will be highlighted, with particular attention to less explored seeds, fermented plant foods and agricultural by-products. Promising future research directions with regards to the application of plant-derived antibacterial hydrolysates and peptides in food preservation, farm animal disease management, and nutraceutical/functional food development will be proposed.
Dysregulated hepatic gluconeogenesis is a hallmark of insulin resistance and type 2 diabetes mellitus (T2DM). Although existing drugs have been proven to improve gluconeogenesis, achieving this objective with functional food is of interest, especially using conjugated linoleic acid (CLA) found in dairy products. Both cis-9, trans-11 (c9,t11) and trans-10, cis-12 (t10,c12) isomers of CLA were tested in human (HepG2) and rat (H4IIE) hepatocytes for their potential effects on gluconeogenesis. The hepatocytes exposed for 24 h with 20 μM of c9,t11-CLA had attenuated the gluconeogenesis in both HepG2 and H4IIE by 62.5% and 80.1%, respectively. In contrast, t10,c12-CLA had no effect. Of note, in HepG2 cells, the exposure of c9,t11-CLA decreased the transcription of gluconeogenic enzymes, cytosolic phosphoenolpyruvate carboxykinase (PCK1) by 87.7%, and glucose-6-phosphatase catalytic subunit (G6PC) by 38.0%, while t10,c12-CLA increased the expression of G6PC, suggesting the isomer-specific effects of CLA on hepatic glucose production. In HepG2, the peroxisome proliferator-activated receptor (PPAR) agonist, rosiglitazone, reduced the glucose production by 72.9%. However, co-administration of c9,t11-CLA and rosiglitazone neither exacerbated nor attenuated the efficacy of rosiglitazone to inhibit glucose production; meanwhile, t10,c12-CLA abrogated the efficacy of rosiglitazone. Paradoxically, PPARγ antagonist GW 9662 also led to 70.2% reduction of glucose production and near undetectable PCK1 expression by abrogating CLA actions. Together, while the precise mechanisms by which CLA isomers modulate hepatic gluconeogenesis directly or via PPAR warrant further investigation, our findings establish that c9,t11-CLA suppresses gluconeogenesis by decreasing PEPCK on hepatocytes.
Matched MeSH terms: Intracellular Signaling Peptides and Proteins/antagonists & inhibitors*; Intracellular Signaling Peptides and Proteins/genetics; Intracellular Signaling Peptides and Proteins/metabolism
The cockle, Anadara granosa, was experimentally exposed to low (0.1 mg/L) and sublethal (1.0 mg/L) doses of copper (Cu) for a period of 24 hrs. Significant increase in Cu concentrations in whole tissues and hepatopancreas compared to control animals were observed. In order to study the effect of copper exposure at molecular levels, a subtractive cDNA library was constructed from the hepatopancreas of cockles exposed to 1.0 mg/L Cu. Screening of the subtractive cDNA library using reverse northern analysis resulted in several differentially expressed genes, including one that codes for metallothionein (MT). The complete coding sequence of the MT gene (designated as AnaMT2) reveals an open reading frame of 234 bp in length that encodes a 77 amino acid polypeptide as revealed by the deduced amino acid composition. Although showing similarities with other molluscan MTs, AnaMT2 can be distinguished by its lower glycine and higher asparagine and proline content. Expression analysis of the AnaMT2 by northern analysis indicated higher mRNA level in cockle exposed to 1.0 mg/L Cu and was undetectable in those treated with 0.1 mg/L. This suggests that AnaMT2 represents a primarily inducible MT not highly expressed under basal conditions.
This is the first report of QQQ-mass spectrometric identification and quantification of the Hb subunits, alpha, beta, delta and gamma globin peptides, derived from enzymatic-digestion of proteins in the early unknown peaks of the Bio-Rad cation-exchange chromatography of haemoglobin. The objectives were to assess the relationship of the quantity of the free alpha, beta, delta and gamma globin chains with the phenotypic diversity of beta-thalassaemias (β-thal). The results demonstrate that the pools of free globin chains in red blood cells were correlating with the severity of the disease in patients with different phenotypes of β-thal. The mechanism and the regulation of synthesis of free globin chains pool in a normal individual and in patients with different β-thal phenotypes could arise from several mechanisms which will require further investigation. The role of the free globin pool in patients with β-thal for development of novel therapeutic approaches based on these potential targets requires further investigation. Pertinent biomarkers improves the diagnosis of the β-thal, especially in low-income countries where they are most common and allows more effective therapeutic intervention leading to more successful therapeutic outcome.
Cellulose nanocrystals (CNC) from mengkuang leaves (Pandanus tectorius) were investigated as potential reinforcement
in poly(vinyl chloride) (PVC) matrix. The surface of CNC was modified with silane coupling agent to improve fillermatrix
adhesion. Solution casting method was used to prepare PVC nanocomposites with various amounts of modified
(SCNC) and unmodified (CNC) nanocrystals. Both SCNC and CNC were examined by Fourier transform infrared (FTIR)
spectroscopy and X-ray diffraction (XRD) which showed that surface chemical modification has occurred. An increase
in tensile strength was observed with the addition of SCNC compared to the CNC. However, the elongation at break of the
nanocomposites was found to decrease with the increase of both fillers loading. An increasing trend was observed in the
tensile modulus with the addition of CNC to the PVC matrix, but decreasing with the addition of SCNC. The morphology
of a fractured surface of nanocomposites showed silane modification reduced the number of voids in the structure of
PVC. The observation indicated the adhesion between the fiber and the matrix had improved upon surface modification
of the nanocrystals with silane.
Matched MeSH terms: Intracellular Signaling Peptides and Proteins
Polyhydroxyalkanoates (PHAs) has been investigated for more than eighty years. Ever since then, the scientists are kept on synthesizing and developing new polymers and application to suit human interests nowadays. The resourcefulness of PHAs has made them a good candidates for the study of their potential in a variety of areas from biomedical/medical fields to food, packaging, textile and household material. In medical field (specifically in tissue engineering application), producing a biocompatible 3-D scaffold with adaptable physical properties are essential. However, to the best of our knowledge, scaffolds from PHB and PHBV with thickness greater than 1 mm have not been produced yet. In this work, PHB and PHBV porous 3-D scaffolds with an improved thickness greater than 4 mm was fabricated using conventional method of solvent-casting particulate-leaching (SCPL). A preliminary assessment on the improved thickness 3-D scaffolds was carried out to examine its pore interconnectivity by using non-invasive color staining method. The vertical cross sections image of the stained scaffolds was analyzed by image analyzer software. This technique was considered simple, fast and cost effective method prior to the usage of super accurate analytical instruments (micro-computed tomography). The results showed that over 80% of the pores have been interconnected with the adjacent pores. Moreover, there was a good correlation between the predicted pore interconnectivity and porosity. These results indicated how well a simple technique can do by giving an overview of the internal morphology of a porous 3-D structure material.
Matched MeSH terms: Intracellular Signaling Peptides and Proteins
The present study investigates heavy metal uptake and protein expression by different vegetables collected from various districts of Khyber Pakhtunkhawa province of Pakistan. Statistical analysis of the data showed that maximum concentration of Cd, Cr, Ni, Zn were found in radish and spinach, respectively, collected from Peshawar. Maximum Pb and Mg accumulation were found in cauliflower and pea at Swat followed by coriander at Haripur and minimum Pb uptake was noticed in radish taken from Nowshehra. Highest Cu uptake was detected in spinach at Nowshehra. Data regarding Cd, Cr, Pb, Cu, Ni, Mg and Zn concentration in water samples gathered from different sites of KPK indicated that maximum concentration of Cd was observed in Swat. Maximum Cr and Cu concentration were measured in water samples from Peshawar while maximum concentration of Pb and Ni were detected in water samples from Haripur. In case of soil samples, maximum Cd, Cr, Mg and Zn uptake was observed in soil sample at Nowshehra. Maximum Pb and Ni concentration was found in soil samples collected from Peshawar. Cu concentration was observed to be the highest in soil at Swat. Protein profile of different vegetables i.e. cauliflower, radish, carrot, turnip, pea, spinach, coriander and garlic sampled across five different sites showed that uptake of Cd, Cr, Pb, Cu, Ni, Mg and Zn by these vegetables caused the expression of numerous polypeptides.
Tilapia is a popular freshwater fish and among the important cultured fish grown worldwide. In this study, fish protein
hydrolysate was produced from tilapia (Oreochromis niloticus) by-product (TB) and tilapia muscle (TM) through enzymatic
hydrolysis using alcalase. The TB and TM protein hydrolysates were evaluated for its characteristics in terms of angiotensin
I-converting enzyme (ACE) inhibition activity, peptide size distribution, and functional properties. Hydrolysis for 1 h for
TB and TM successfully produced low molecular weight peptides (<14.2kDa) with the highest ACE inhibitory activities.
The findings also demonstrated that both samples have high nitrogen solubility (>80% at pH2-9) and good emulsifying,
water and oil holding capacities. The study indicated that tilapia protein hydrolysates have the potential to be used as
functional food products.
Chitosan/polyvinyl alcohol (Chitosan/PVA) blended film was prepared by direct blend process and solution casting methods.
In order to reduce the swelling ratio and enhance the chemical and mechanical stability, Chitosan/PVA film was crosslinked
with glutaraldehyde in order to produce Chitosan-g-PVA. Bovine serum albumin (BSA) was used as a model protein
to incorporate into the Chitosan-g-PVA. The chemical structure and morphological characteristics of films were studied
by FT-IR and scanning electron microscopy (SEM). Mechanical and physical properties of blended films such as tensile
properties in the dry and wet states, water uptake and water contact angle measurement were characterized. Blending
PVA and chitosan improved strength and flexibility of the films. Crosslinking with glutaraldehyde further improves the
tensile strength and decrease the hydrophilicity of films. BSA immobilized on the Chitosan-g-PVA film was calculated as
BSA encapsulation efficiency.
Matched MeSH terms: Intracellular Signaling Peptides and Proteins
This is the first report of quadrupole time-of-flight (Q-TOF) mass spectrometric identification of the hemoglobin (Hb) subunits, α, β, δ and γ peptides, derived from enzymatic-digestion of proteins in the early unknown peaks of the cation exchange chromatography of Hb. The objectives were to identify the unknown high performance liquid chromatography (HPLC) peaks in healthy subjects and in patients with β-thalassemia (β-thal). The results demonstrate the existence of pools of free globin chains in red blood cells (RBCs). The α-, β-, δ- and γ-globin peptides were identified in the unknown HPLC peaks. The quantification and role of the free globin pool in patients with β-thal requires further investigation. Identification of all types of Hb subunits in the retention time (RT) before 1 min. suggests that altered Hbs is the nature of these fast-eluting peaks. Relevancy of thalassemias to the protein-aggregation disorders will require review of the role of free globin in the pathology of the disease.
Objective: The assay for anti-cyclic citrullinated peptide (anti-CCP) antibody is a recent test of interest due to low diagnostic specificity of rheumatoid factor in the diagnosis of rheumatoid arthritis. To determine the sensitivity and specificity of anti-CCP antibodies in rheumatoid arthritis patients using American College of Rheumatology (ACR) criteria as a gold standard.
Design: A cross sectional study was conducted from January 2008 to December 2008 on 100 patients with rheumatoid arthritis and 153 patients with arthritis or arthralgia but not fulfilling ACR criteria for rheumatoid arthritis.
Materials and Methods: Serum from each subject was tested for anti-CCP antibodies and IgM rheumatoid factor (IgM RF) by an enzyme linked immunosorbent assay (ELISA). Sensitivity and specificity of the tests were evaluated using the ACR criteria as the gold standard. Data was analyzed using SPSS version 12.0 software. The association between the two categorical variables were tested using chi-square test. The differences between the two groups were tested using independent t-test. Results: The sensitivity of anti-CCP antibodies was 71% with 94.8% of specificity. For rheumatoid factor the sensitivity was 85% and the specificity was 74.5%. Positive predictive value for anti-CCP antibodies was 89.9% whereas for rheumatoid factor it was 68.5%. The sensitivity of ACR criteria with inclusion of anti- CCP antibodies was 100% and specificity of 94.8% (95%CI: 91.3%, 98.2%).
Conclusion: Anti-CCP antibody has a higher diagnostic accuracy compared to the RF. Anti-CCP antibody is a useful laboratory marker to support the diagnosis of rheumatoid arthritis and to differentiate patients with rheumatoid arthritis from rheumatoid arthritis-like symptoms. Therefore, it is suggested that anti-CCP antibodies should be included as another laboratory supportive criterium besides RF test in ACR criteria in the diagnosis of RA.
Study site: family medicine clinic, rheumatology clinic and immunology laboratory of Hospital Universiti Sains Malaysia (HUSM), Kubang Kerian, Kelantan, Malaysia