Background: Anti-cyclic citrullinated peptide autoantibodies (anti-CCP) is a new serological test for the diagnosis of rheumatoid arthritis (RA). It is an enzyme immunoassay (EIA) for the detection of antibodies directed toward citrullinated peptides. Studies show this test has an improved diagnostic value compared to rheumatoid factor (RF). Objective: To determine the sensitivity and specificity of anti-CCP in patients with rheumatoid arthritis and other rheumatic diseases. Method: 227 serum samples for rheumatology clinics (Putrajaya, Taiping, and Ipoh Hospital) were tested for the presence of anti-CCP and rheumatoid factor (RF). These included 171 patients diagnosed with RA and 56 from other rheumatic diseases. Patient demographic data, clinical diagnosis, radiographic information and other laboratory data were obtained from the patients' clinical notes. Results: Anti-CCP antibodies were detected in 76.6% (131/171) patients with RA and 17.9% (10/56) patients with other arthritis. The sensitivity and specificity of anti-CCP reactivity at the optimal cut off values were 66.1% and 87.5% respectively. The sensitivity of anti-CCP was higher than that for RF (41.8%). However, the presence of either anti-CCP or RF improved the sensitivity to 76.2%. Conclusion: The detection of anti-CCP alone maybe useful in the diagnosis of RA. However, when used concomitantly with RF, it can improve the diagnostic ability significantly.
Using radiolabelled peptides that bind, with high affinity and specificity, to receptors on tumour cells is one of the most promising fields in modern molecular imaging and targeted radionuclide therapy (1). In the emergence of molecular imaging and nuclear medicine diagnosis and therapy, albeit theranostic, radiolabelled peptides have become vital tools for in vivo visualisation and monitoring physiological and biochemical processes on molecular and cellular levels (2). This approach may benefit patients in the era of personalised medicine.
Antibacterial peptides or bacteriocins produced by many strains of lactic acid bacteria have been used as food preservatives for many years without any known adverse effects. Bacteriocin titres can be modified by altering the physiological and nutritional factors of the producing bacterium to improve the production in terms of yield and productivity. The effects of culture conditions (initial pH, inoculum age and inoculum size) and medium compositions (organic and inorganic nitrogen sources; carbon sources) were assessed for the production of bacteriocin-like inhibitory substances (BLIS) by Lactococcus lactis Gh1 in shake flask cultures. An inoculum of the mid-exponential phase culture at 1% (v/v) was the optimal age and size, while initial pH of culture media at alkaline and acidic state did not show a significant impact on BLIS secretion. Organic nitrogen sources were more favourable for BLIS production compared to inorganic sources. Production of BLIS by L. lactis Gh1 in soytone was 1.28-times higher as compared to that of organic nitrogen sources ((NH4)2SO4). The highest cell concentration (XmX = 0.69 ± 0.026 g·L-1) and specific growth rate (μmax = 0.14 h-1) were also observed in cultivation using soytone. By replacing carbon sources with fructose, BLIS production was increased up to 34.94% compared to BHI medium, which gave the biomass cell concentration and specific growth rate of 0.66 ± 0.002 g·L-1 and 0.11 h-1, respectively. It can be concluded that the fermentation factors have pronounced influences on the growth of L. lactis Gh1 and BLIS production. Results from this study could be used for subsequent application in process design and optimisation for improving BLIS production by L. lactis Gh1 at larger scale.
Matrix metalloproteinases (MMPs) are enzymes that can degrade collagen in hybrid layer and reduce the longevity of adhesive restorations. As scientific understanding of the MMPs has advanced, useful strategies focusing on preventing these enzymes' actions by MMP inhibitors have quickly developed in many medical fields. However, in restorative dentistry, it is still not well established. This paper is an overview of the strategies to inhibit MMPs that can achieve a long-lasting material-tooth adhesion. Literature search was performed comprehensively using the electronic databases: PubMed, ScienceDirect and Scopus including articles from May 2007 to December 2019 and the main search terms were "matrix metalloproteinases", "collagen", and "dentin" and "hybrid layer". MMPs typical structure consists of several distinct domains. MMP inhibitors can be divided into 2 main groups: synthetic (synthetic-peptides, non-peptide molecules and compounds, tetracyclines, metallic ions, and others) and natural bioactive inhibitors mainly flavonoids. Selective inhibitors of MMPs promise to be the future for specific targeting of preventing dentin proteolysis. The knowledge about MMPs functionality should be considered to synthesize drugs capable to efficiently and selectively block MMPs chemical routes targeting their inactivation in order to overcome the current limitations of the therapeutic use of MMPs inhibitors, i.e., easy clinical application and long-lasting effect.
Cyclopolypeptides are among the most predominant biomolecules in nature, especially those derived from plant seeds. This category of compounds has gained extraordinary attention due to remarkable variety of structures and valuable biofunctions. These congeners display enormous variation in terms of both structure and function and are the most significant biomolecules due to their widespread bioproperties. The estrogenic activity, immunosuppressive activity, cytotoxicity, vasorelaxant activity, and other properties possessed by cyclic peptides from seeds of plants make these congeners attractive leads for the drug discovery process. The current study covers the important structural features, structure-activity relationship, synthesis methods, and bioproperties of plant seeds-originated bioactive peptides from Vaccaria segetalis, Linum usitatissimum, and Goniothalamus leiocarpus, which may prove vital for the development of novel therapeutics based on a peptide skeleton.
Haruan or Channa striatus is source of protein that is widely consumed in the region and its extract
is well known for having medical values. It is of great advantage if this product could be taken
orally rather than by injection because the oral route of drug delivery is still preferred by the vast
majority of patients. However protein and peptides can be denatured or degraded by the acidic pH
of the stomach and the presence of endogenous enzymes. In order to protect or prevent digestion
and degradation of the protein in the stomach and to ensure the protein reaches the gastro
intestinal (GI) tract, Carboxymethyl Starch (CMS) nanogel system was developed using electron
irradiation method. However stability of HTE during the irradiation process needed to be studied
before being developed further. In this study, the HTE was irradiated using electron beams. Its
stability was analysed in terms of physical and chemical changes by looking at colour difference,
melting point by using Differential Scanning Calorimetry (DSC) and molecular bonds by using
Fourier Transform Infrared (FTIR) respectively. The results of this study were that no apparent
colour difference was observed with HTE before and after irradiation. These observations were
supported by the FTIR and DSC results that showed that there were no changes in molecular bonds
and melting point, compared between no irradiation and irradiation HTE during electron
irradiation up to 10 kGy. Statistically the test showed no significant difference at p < 0.005 between
melting temperatures.
Threadfin bream (Nemipterus japonicas) muscle was hydrolysed using protease extracted from
bilimbi (Averrhoa bilimbi L.) fruit. This study was performed in order to compare the efficiency of bilimbi protease in producing threadfin bream protein hydrolysate with the commercial protease; alcalase 2.4 L. Initially, protease was extracted and then purified using 40% ammonium sulfate precipitation method. The proteolytic activity of the crude extract and purified protease was determined. Precipitation using 40% ammonium sulfate resulted in bilimbi protease specific activity of 2.36 U/mg and 23.13% recovery. Threadfin bream hydrolysate was prepared based on the pH-stat method by hydrolysis for 2 hrs. Hydrolysis using bilimbi protease produced 34.76% degree of hydrolysis (DH) and 3.75% yield while hydrolysis using alcalase resulted in 86.6% DH with 22.78% yield. Alcalase hydrolysate showed higher solubility than bilimbi protease hydrolysate at pH 7 with 70.87 and 32.16% solubility, respectively. Results also showed that protein content of threadfin bream hydrolysate produced using alcalase was higher (86.86%) than those produced using bilimbi protease (22.12%). However, both hydrolysates showed low moisture content between 3.93 to 7.00%. The molecular weight distribution analysis using SDS–PAGE indicated the distribution of smaller peptides especially in alcalase hydrolysate. Overall, the results showed that alcalase is more efficient enzyme choice than bilimbi protease for preparing threadfin bream hydrolysates. However, both hydrolysates could play an important role thus contribute to the food industry.
The physicochemical properties of silver catfish frame hydrolysate powder at three different degree of hydrolysis, DH43%, DH 55% and DH 68% were studied. The hydrolysates powder were obtained by hydrolysis using Alcalase®, centrifugation and spray drying of the supernatant. The study found that preparation of these hydrolysates affected the protein, ash and fat content as well as amino acid composition. As for essential amino acids, their values were generally considered as adequate as compared to the suggested essential amino acids profile of FAO/WHO. The results showed that SFHs were rich in lysine and glutamate. Hydrolysate at DH 68% exhibited better peptide solubility and water holding capacity. As degree of hydrolysis increased, emulsifying capacity and foaming capacity of the hydrolysate decreased. It was also found that the lightness in hydrolysate powder decreased with increase in degree of hydrolysis. This study shows that silver catfish frame hydrolysate has good solubility, good foaming properties and light colour profile, thus having high potential as food ingredient.
Pepsin-solubilized collagen (PSC) was extracted from the body wall of crown-of-thorns starfish (COTS) (Acanthaster planci) using pepsin digestion in 0.5 M acetic acid. The electrophoretic pattern of PSC showed that it consisted of two α chains (α1 and α2 chains). In addition, the peptide mapping showed that there were some differences in peptide patterns among PSC, calf skin collagen and salmon skin collagen. This suggested that the primary structure of the PSC was different from calf skin collagen and salmon skin collagen. Furthermore, Fourier transform infrared spectroscopy (FTIR) investigation showed the existence of triple helical structure in PSC, suggesting that the triple helical structure was well preserved during the extraction of collagen from COTS. The denaturation temperature of PSC was 33.0°C, which was comparable with mammalian collagen. In addition, the amino acid composition analysis showed that the imino acid content of PSC was similar to mammalian collagen but it was higher than other marine collagen. This suggested that the imino acid content might contributed the denaturation temperature of collagen. The results in this study suggest that PSC from the underutilized COTS has potential to be exploited in various commercial applications.
Milk-derived bioactive peptides have been identified as potential ingredients of health-promoting functional foods. These bioactive peptides are targeted at diet-related chronic diseases especially the non-communicable diseases viz., obesity, cardiovascular diseases and diabetes. Peptides derived from the milk of cow, goat, sheep, buffalo and camel exert multifunctional properties, including anti-microbial, immune modulatory, anti-oxidant, inhibitory effect on enzymes, anti-thrombotic, and antagonistic activities against various toxic agents. Majority of those regulate immunological, gastrointestinal, hormonal and neurological responses, thereby playing a vital role in the prevention of cancer, osteoporosis, hypertension and other disorders as discussed in this review. For the commercial production of such novel bioactive peptides large scale technologies based on membrane separation and ion exchange chromatography methods have been developed. Separation and identification of those peptides and their pharmacodynamic parameters are necessary to transfer their potent functional properties into food applications. The present review summarizes the preliminary classes of bioactive milk-derived peptides along with their physiological functions, general characteristics and potential applications in health-care.
Ebola virus (EBOV) is one of the lethal viruses, causing more than 24 epidemic outbreaks to date. Despite having available molecular knowledge of this virus, no definite vaccine or other remedial agents have been developed yet for the management and avoidance of EBOV infections in humans. Disclosing this, the present study described an epitope-based peptide vaccine against EBOV, using a combination of B-cell and T-cell epitope predictions, followed by molecular docking and molecular dynamics simulation approach. Here, protein sequences of all glycoproteins of EBOV were collected and examined via in silico methods to determine the most immunogenic protein. From the identified antigenic protein, the peptide region ranging from 186 to 220 and the sequence HKEGAFFLY from the positions of 154-162 were considered the most potential B-cell and T-cell epitopes, correspondingly. Moreover, this peptide (HKEGAFFLY) interacted with HLA-A*32:15 with the highest binding energy and stability, and also a good conservancy of 83.85% with maximum population coverage. The results imply that the designed epitopes could manifest vigorous enduring defensive immunity against EBOV.
Microinjection is a powerful tool to deliver various substances, such as nucleic acids, proteins, peptides, RNA, and synthetic molecules into mammalian cells mechanically. Through microinjection, a controlled amount of protein can be delivered into the target cells to elucidate the specific functional
effects in vitro. In this study, a series of protein microinjection optimization was performed in human breast cancer cells. The presence of Maltose Binding Protein (MBP) was microscopically monitored through indirect immunofluorescence assay. The optimization experimentation gave a high success rate when MBP protein was used at the minimum concentration of 1.5 mg/ml and at the injection pressures of 50 and 70 hPa. The average success rate of injections was 49.2±4.15% and 50.8±4.6%, while the average cell survivability was 50.98±4.67% and 49.72±5.48% at 50 and 70 hPa, respectively. The optimization of the MBP concentration and injection pressures successfully allowed an efficient delivery of precise protein dosage into breast cancer cells without any adverse effect. This microinjection optimization can be a practical guideline in any downstream applications of protein functional work.
Gelatin is widely used in food and pharmaceutical products. However, the addition of gelatin especially in food products becomes a controversial issue among Muslims due to its animal origin. Thus, the present study was aimed to detect and differentiate the origin of gelatin added in processed foods using a combination method of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA). Porcine gelatin had exhibited 11 prominent polypeptides compared to bovine gelatin with 2 prominent polypeptides. Polypeptides of both gelatin sources at molecular weight ranged from 53 to 220 kDa can be used to differentiate between porcine and bovine gelatins using PCA. The efficiency in extracting gelatin from processed foods by different solutions was also evaluated. Extraction of gelatin in processed foods by cold acetone and deionised water had exhibited a similar polypeptide patterns, suggesting both solutions are suitable. The study indicated that approach of a simple gelatin extraction combined with SDS-PAGE and PCA, may provide robust information for gelatin species differentiation of processed foods.
Different heat treatments, (1) chilled, 4°C (2) boiled at 100°C for 30 min and (3) autoclaved at 121°C at 15 psi for 20 min were employed on goat meat to mimic domestic and industrial cooking. The effects on intensity of actin proteins was observed using two-dimensional gel electrophoresis where significant differences (p>0.05) were observed in the spot intensity between chilled and boiled samples, similarly in chilled and autoclaved samples. However, no significant difference was observed between boiled and autoclaved samples. The slight changes observed in the cooking of meat confirmed that actin protein is susceptible to denaturation cause by heat. MALDI-TOF/TOF analysis revealed the peptide-mass fingerprint between positions 21 – 374 that not affected by heat treatment. Peptides from this position merit the candidature of actin as putative thermostable marker for detecting goat meat (chevon) in food product.
The amino-acid composition, 2, 2-Diphenyl-1-picryhydrazyl (DPPH) radical-scavenging activity, and peptide patterns of tilapia protein hydrolysates produced by the enzymatic hydrolysis of Alcalase (AH), Flavourzyme (FH) and Protamex (PH) for 5h using pH-stat method were studied. The ratio of essential amino acids to non-essential amino acids increased after hydrolysis in all samples; however, no significant differences among them were observed. AH had a highest (P < 0.05) DPPH radical-scavenging activity, but no significant difference in the DPPH between FH and PH was observed. SDS-PAGE patterns for all the hydrolysates showed significant (P < 0.05) reduction in the number and the intensity of the bands with increasing time of hydrolysis. Flavourzyme showed the lowest rate of hydrolytic activity towards the tilapia mince.
The sea cucumber (Stichopus vastus) is an underutilized species, as most of its parts, including the integument (high collagen content) are thrown away during processing. The aim of this study was to investigate the effects of different hydrolysis conditions (substrate to enzyme ratio (S/E), reaction temperature, and hydrolysis time) on the angiotensin I converting enzyme (ACE) inhibitory and radical scavenging (RSc) activities of the hydrolysates produced from trypsin hydrolysis of S. vastus collagen. Optimal conditions predicted by Box-Behnken Design modelling for producing ACE inhibitory and RSc hydrolysates were found to be S/E ratio (15), reaction temperature (55°C), and hydrolysis time (1 h). Under optimal conditions, ACE inhibitory and RSc activities were estimated to be as high as 67.8% and 77.9%, respectively. Besides, some novel bioactive peptides were identified through mass spectrometry analysis. These results indicate that S. vastus hydrolysates might be used as a functional ingredient in food and nutraceutical products.
The aim was to examine the protein profiles of whole and parotid saliva using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and MALDI-TOF mass spectrometry. The banding patterns of proteins exhibited by the unstimulated whole saliva samples on the gel remained quite constant but the intensity of the protein bands were slightly different from one sample to another. Comparison of the protein profiles of unstimulated whole saliva and stimulated parotid saliva showed almost similar banding pattern. The exception is the presence of a pink protein band in the 65-67 kD region in the stimulated parotid saliva samples which was also observed in the unstimulated whole saliva sample contributed by a cerebral palsy patient. Analysis of the saliva samples using MALDI-TOF mass spectrometry also revealed that the stimulated parotid saliva samples exhibited some peaks that were in the same region as those for the unstimulated whole saliva sample of the cerebral palsy subject. This may imply that there is ineffective control of the parotid secretion in cerebral palsy subject under unstimulated condition. The SDS-PAGE and MALDI-TOF analyses may provide more information on the profiles of the salivary proteins which could be beneficial in the diagnosis of salivary gland dysfunction.
Matched MeSH terms: Salivary Proteins and Peptides
Natural peptides in insect vectors played an important role in the control of
pathogens. Musca domestica Linnaeus and Chrysomya megacephala Fabricius were
two species of local fly chosen to detect presence of antimicrobial peptide substance.
The screening of the antimicrobial activity was carried using a spectrophotometric
method. Results were obtained much quicker and less laborious. The results showed
larva hemolymph of M. domestica lysed Bacillus subtilis and two Gram negatives,
Escherichia coli and Pseudomona. aeruginosa. The pupae hemolymph only lysed E.
coli. Whereas, the hemolymph of C. megacephala larva showed bactericidal effect
against both of the Gram positives tested, i.e. B. subtilis and Staph. aureus. and no
effect was against the Gram negatives. The pupa showed lytic activity against Staph.
aureus and P. aeruginosa. As a conclusion, the larva and pupa hemolymph of M.
domestica and C. megacephala demonstrated antibacterial activity. However, larva
hemolymph of M. domestica and C. megacephala has broader antibacterial activity
against both Gram positive and negative bacteria.
Introduction: During the last three decades hematopoietic stem cell transplantation (HSCT) has become a well-established treatment for many hematologic malignancies. The most important limitation for HSC transplantation is the low number of hematopoietic stem cells (HSC) that can lead to delayed engraftment or graft failures. Numerous attempts have been made to improve in vitro HSC expansion via optimization of various methods such as isolation techniques, supplementing with growth factors, utilizing stromal cells as feeder layer and other culture conditions. Objective: This project is aimed to decipher the efficiency of an isolation technique and retrieval of culture expanded HSC from feeder layer using two different harvesting methods. Materials and Methods: Hematopoietic stem cells from human umbilical cord blood were isolated via MACS mediated CD34+ double sorting. Then, the cells were cultured onto MSC feeder layer for 3 and 5 days. Culture expanded cells were harvested using two different harvesting method namely cell aspiration and trypsinization methods. Hematopoietic stem cell expansion index were calculated based on harvesting methods for each time point. Results: The numbers of HSC isolated from human umbilical cord blood were 1.64 x 106 and 1.20 x106 cells at single and double sortings respectively. Although the number of sorted cells diminished at the second sorting yet the yield of CD34+ purity has increased from 43.73% at single sorting to 81.40% at double sorting. Employing the trypsinization method, the HSC harvested from feeder layer showed a significant increase in expansion index (EI) as compared to the cell aspiration harvesting method (p≤ 0.05). However, the purity of CD34+ HSC was found higher when the cells were harvested using aspiration method (82.43%) as compared to the trypsinization method (74.13%). Conclusion: A pure population of CD34+ HSC can be retrieved when the cells were double sorted using MACS and expanded in culture after being harvested using cell aspiration method.
Matched MeSH terms: Intercellular Signaling Peptides and Proteins
Safe immunostimulants (adjuvants) are essential for the development of highly potent peptide-based vaccines. This study examined for the first time whether fluorinated lipids could stimulate humoral immunity in vivo when conjugated to peptide antigen. The impact of fluorination on humoral immunity was tested using a library of peptide-based vaccine candidates against the group A streptococcus (GAS). The fluorinated constructs stimulated similar mouse IgG titers to those elicited by complete Freund's adjuvant (CFA) and were higher than those produced in mice that received the nonfluorinated constructs.