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Heavy and irresponsible use of antibiotics in the last century has put selection pressure on the microbes to evolve even faster and develop more resilient strains. In the confrontation with such sometimes called "superbugs", the search for new sources of biochemical antibiotics seems to have reached the limit. In the last two decades, bioactive antimicrobial peptides (AMPs), which are polypeptide chains with less than 100 amino acids, have attracted the attention of many in the control of microbial pathogens, more than the other types of antibiotics. AMPs are groups of components involved in the immune response of many living organisms, and have come to light as new frontiers in fighting with microbes. AMPs are generally produced in minute amounts within organisms; therefore, to address the market, they have to be either produced on a large scale through recombinant DNA technology or to be synthesized via chemical methods. Here, heterologous expression of AMPs within bacterial, fungal, yeast, plants, and insect cells, and points that need to be considered towards their industrialization will be reviewed. Sources of peptide production and their applications. Some AMPs directly extracted from natural sources, some of them are chemically synthesized either using liquid or solid phase peptides synthesis, and for large scale production, recombinant expression using heterologous expression systems have been used. [ABSTRACT FROM AUTHOR]

Rapid and label-free detection of very low concentrations of biomarkers in disease diagnosis or therapeutic drug monitoring is essential to prevent disease progression in Point of Care Testing. For this purpose, we propose a multi-purpose optical Bio-Micro-Electro-Mechanical-System (BioMEMS) sensing platform which can precisely measure very small changes of biomolecules concentrations in plasma-like buffer samples. This is realized by the development of an interferometric detection method on highly sensitive MEMS transducers (cantilevers). This approach facilitates the precise analysis of the obtained results to determine the analyte type and its concentrations. Furthermore, the proposed multi-purpose platform can be used for a wide range of biological assessments in various concentration levels by the use of an appropriate bioreceptor and the control of its coating density on the cantilever surface. In this study, the present system is prepared for the identification of digoxin medication in its therapeutic window for therapeutic drug monitoring as a case study. The experimental results represent the repeatability and stability of the proposed device as well as its capability to detect the analytes in less than eight minutes for all samples. In addition, according to the experiments carried out for very low concentrations of digoxin in plasma-like buffer, the detection limit of LOD = 300 fM and the maximum sensitivity of S = 5.5 × 1012 AU/M are achieved for the implemented biosensor. The present ultrasensitive multi-purpose BioMEMS sensor can be a fully-integrated, cost-effective device to precisely analyze various biomarker concentrations for various biomedical applications. [ABSTRACT FROM AUTHOR]

Background: Current approaches used to overcome microbial infections are becoming inefficient due to the overuse or misuse of antibiotics. Antimicrobial peptides (AMPs) are one of the most promising substitutional candidates for commercial antibiotics. Methods and Results: The publications in the field of in silico design of AMPs with focus on the wound-healing AMPs were searched though SCOPUS and PubMed. Through publications, it was reported that a number of AMPs approved for clinical use have also showed efficient wound-healing activity. Todays, the design and production of synthetic types of AMPs have attracted attention in order to expand their applications and to cope with their limitations and adverse effects. In this paper, the currently published researches in the field of AMPs and their wound-healing features were summarized and the strategies used in AMPs design and development have been reviewed. Moreover, different databases and online servers used in this regard were summarized. Conclusion: Design and development of active AMPs, especially those with wound-healing activity, can be performed using bioinformatics servers and software, regarding AMPs structure-activity relationships. [ABSTRACT FROM AUTHOR]

Aptamers can be regarded as efficient substitutes for monoclonal antibodies in many diagnostic and therapeutic applications. Due to the tedious and prohibitive nature of SELEX (systematic evolution of ligands by exponential enrichment), the in silico methods have been developed to improve the enrichment processes rate. However, the majority of these methods did not show any effort in designing novel aptamers. Moreover, some target proteins may have not any binding RNA candidates in nature and a reductive mechanism is needed to generate novel aptamer pools among enormous possible combinations of nucleotide acids to be examined in vitro. We have applied a genetic algorithm (GA) with an embedded binding predictor fitness function to in silico design of RNA aptamers. As a case study of this research, all steps were accomplished to generate an aptamer pool against aminopeptidase N (CD13) biomarker. First, the model was developed based on sequential and structural features of known RNA–protein complexes. Then, utilizing RNA sequences involved in complexes with positive prediction results, as the first-generation, novel aptamers were designed and top-ranked sequences were selected. A 76-mer aptamer was identified with the highest fitness value with a 3 to 6 time higher score than parent oligonucleotides. The reliability of obtained sequences was confirmed utilizing docking and molecular dynamic simulation. The proposed method provides an important simplified contribution to the oligonucleotide–aptamer design process. Also, it can be an underlying ground to design novel aptamers against a wide range of biomarkers. [ABSTRACT FROM AUTHOR]

Microalgae and cyanobacteria are the potentially valuable source of bioactive compounds applied in the various industries and human usage in different fields of pharmaceutical, nutraceutical, and cosmetic disciplines. One of the interesting aspects is their application as the anti-inflammatory agents for treatment of inflammation related mal-conditions. Natural compounds are of great importance in the treatment of inflammations to reduce the reaction of immune system against pathogens, toxic compounds and damaged cells. A wide range of different metabolites with various chemical structures, including small molecules and peptides and proteins, polysaccharides, fatty acids and their derivatives have been found in microalgae and cyanobacteria which have anti-inflammatory activity. In this review, we summarized different metabolites with anti-inflammatory activity that were extracted from these microorganisms and their mechanisms. The bioactive compounds from microalgae and cyanobacteria have exhibited anti-inflammatory activity through different mechanisms acting intra- or extra- cellularly. So, they could be considered as promising anti-inflammatory agents in treatment of related diseases. [ABSTRACT FROM AUTHOR]

Recently, much attention has been focused on the use of miRNAs in cancer treatment. The role of proto-oncogene Janus kinase-2 (JAK-2) in proliferation and survival of gastric cancer has been previously documented. The aim of this study was to evaluate the effect of a chimera consisted of nucleolin specific aptamer (NCL-Apt) and miRNA let-7d on JAK2 expression level and activity in gastric cancer cells. NCL-Apt-miRNA let-7d chimera was prepared by two methods. Gastric cancer (MKN-45) cell line and control cell line of human dermal fibroblast (HDF) were treated with the chimera and the changes in JAK2 expression and activity were determined using real-time PCR and ELISA techniques, respectively. In MKN-45 cells, the chimera caused significant decrease in JAK2 expression level and activity compared to the aptamer alone and miRNA mimic negative control. Nevertheless, transfected miRNA let-7d showed remarkable reduction in the expression level of JAK2 in comparison with control state in both MKN-45 and HDF, confirmed unspecific effect of let-7d on normal and cancerous cells. With regard to the synergic effect of this chimera on JAK2 activity, it might be viewed as a therapeutic candidate in gastric cancer. However, further studies are warranted to prove it. [ABSTRACT FROM AUTHOR]

Background: In the pharmaceutical industry, hard- and soft-shelled capsules are typically made from gelatin, commonly derived from bovine and porcine sources. To ensure that pharmaceutical products comply with halal regulations in Muslim countries (no porcine products allowed), development of a valid, reliable, quick, and most importantly, cost-effective tests are of utmost importance. Methods: We developed a species-specific duplex polymerase chain reaction (PCR) assay targeting 149 bp porcine and 271 bp bovine mitochondrial DNA (mtDNA) to simultaneously detect both porcine and bovine DNA (in one reaction at the same time) in gelatin. Some additional simplex PCR tests (targeting 126 bp bovine and 212 bp porcine mtDNA) and real-time PCR using a commercially available kit (for identification of porcine DNA) were used to verify the selectivity and sensitivity of our duplex PCR. After optimization of DNA extraction and PCR methods, hard/soft pharmaceutical gelatin capsules (containing drug) were tested for the presence of porcine and/or bovine DNA. Results: Duplex PCR detected the presence of as little as 0.1% porcine DNA, which was more accurate than the commercially available kit. Of all gelatin capsules tested (n = 24), 50% contained porcine DNA (pure porcine gelatin alone or in combination with bovine gelatin). Conclusions: Duplex PCR presents an easy-to-follow, quick, low-cost and reliable method to simultaneously detect porcine and bovine DNAs (>100 bp) in minute amounts in highly processed gelatin-containing pharmaceutical products (with a 0.1% sensitivity for porcine DNA) which may be used for halal authentication [ABSTRACT FROM AUTHOR]

Aptamers are single stranded oligonucleotides, comparable to monoclonal antibodies (mAbs) in selectivity and affinity and have significant strategic properties in design, development and applications more than mAbs. Ease of design and development, simple chemical modification and the attachment of functional groups, easily handling and more adaptability with analytical methods, small size and adaptation with nanostructures are the valuable characteristics of aptamers in comparison to large protein based ligands. Among a broad range of targets that their specific aptamers developed, proteins and peptides have significant position according to the number of related studies performed so far. Since proteins control many of important physiological and pathological incidents in the living organisms, particularly human beings and because of the benefits of aptamers in clinical and analytical applications, aptamer related technologies in the field of proteins and peptides are under progress, exclusively. Currently, there is only one FDA approved therapeutic aptamer in the pharmaceutical market, which is specific to vascular endothelial growth factor and is prescribed for age related macular degenerative disease. Additionally, there are several aptamers in the different phases of clinical trials. Almost all of these aptamers are specific to clinically important peptide or protein targets. In addition, the application of protein specific aptamers in the design and development of targeted drug delivery systems and diagnostic biosensors is another intersting field of aptamer technology. In this review, significant efforts related to development and applications of aptamer technologies in proteins and peptides sciences were considered to emphasis on the importance of aptamers in medicinal and clinical applications. [ABSTRACT FROM AUTHOR]