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1.
Int J Biol Macromol ; 272(Pt 1): 132869, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38838895

ABSTRACT

Repurposing discarded cells stands as a groundbreaking paradigm shift in sustainable biotechnology, with profound implications across diverse industrial sectors. Our study proposes a transformative concept by harnessing histone proteins from discarded CHO cells to produce bioactive peptides. We systematically isolated and hydrolyzed histones using Trypsin and Neutrase enzymes, optimizing reaction conditions. Ultrafiltration yielded distinct peptide fractions (<3 kDa and 3-10 kDa), which we analyzed for DPP-IV inhibition, antioxidant potential, and other activities. Furthermore, LC-Q-TOF-MS analysis and in silico tools unveiled the structural composition of bioactive peptides within these fractions. Three peptide sequences with high bioactivity potential were identified: KLPFQR, VNRFLR, and LSSCAPVFL. Our findings demonstrated exceptional DPP-IV inhibition, potent antioxidant effects, and effective anti-lipid peroxidation activities, surpassing reference compounds. Hemolytic activity assessment indicated promising biocompatibility, enhancing therapeutic application prospects. Pioneering the strategic repurposing of discarded cells, this research addresses cost-efficiency in cell-based studies and promotes sustainable use of biological resources across sectors. This novel approach offers an efficient, eco-friendly method for bioactive molecule procurement and resource management, revolutionizing cell culture studies and biotechnological applications.


Subject(s)
Biotechnology , Cricetulus , Peptides , CHO Cells , Animals , Peptides/chemistry , Peptides/pharmacology , Biotechnology/methods , Antioxidants/pharmacology , Antioxidants/chemistry , Histones/metabolism , Histones/chemistry , Amino Acid Sequence , Hemolysis/drug effects
2.
Biosensors (Basel) ; 13(3)2023 Mar 18.
Article in English | MEDLINE | ID: mdl-36979612

ABSTRACT

Minimally invasive approaches for cancer diagnosis are an integral step in the quest to improve cancer survival. Liquid biopsies such as blood samples are matrices explored to extract valuable information about the tumor and its state through various indicators, such as proteins, peptides, tumor DNA, or circulating tumor cells. Although these markers are scarce, making their isolation and detection in complex matrices challenging, the development in polymer chemistry producing interesting structures, including molecularly imprinted polymers, branched polymers, nanopolymer composites, and hybrids, allowed the development of enhanced platforms with impressive performance for liquid biopsies analysis. This review describes the latest advances and developments in polymer synthesis and their application for minimally invasive cancer diagnosis. The polymer structures improve the operational performances of biosensors through various processes, such as increased affinity for enhanced sensitivity, improved binding, and avoidance of non-specific interactions for enhanced specificity. Furthermore, polymer-based materials can be a tremendous help in signal amplification of usually low-concentrated targets in the sample. The pros and cons of these materials, how the synthesis process affects their performance, and the device applications for liquid biopsies diagnosis will be critically reviewed to show the essentiality of this technology in oncology and clinical biomedicine.


Subject(s)
Neoplasms , Humans , Liquid Biopsy , Neoplasms/diagnosis , Neoplasms/pathology , DNA , Polymers/chemistry , Proteins
3.
Phytochem Anal ; 33(7): 1058-1067, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35795911

ABSTRACT

INTRODUCTION: The chronicity of advanced glycation end-products (AGEs) imparts various damages resulting in metabolic dysfunction and diseases involving inflammation and oxidative stress. The use of plant extracts is of high interest in complementary medicine. Yet, extracts are multicomponent mixtures, and difficult to pinpoint their exact mechanism. OBJECTIVES: We hypothesise that network pharmacology and bioinformatics can help experimental findings depict the exact active components and mechanism of action by which they induce their effects. Additionally, the toxicity and variability can be lowered and standardised with proper encapsulation methods. METHODOLOGY: Here, we propose the formulation of phytoniosomes encapsulating two Artemisia species (Artemisia dracunculus and Artemisia absinthium) to mitigate AGEs and their induced cell redox dysregulation in the liver. Extracts from different solvents were identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS/MS). Phytoniosomes were explored for their anti-glycating effect and modulation of AGE-induced damages in THLE-2 liver cells. Network pharmacology tools were used to identify possible targets and signalling pathways implicated. RESULTS: Data demonstrated that A. absinthium phytoniosomes had a significant anti-AGE effect comparable to reference molecules and higher than A. dracunculus. They were able to restore cell dysfunction through the restoration of tumour necrosis alpha (TNF-α), interleukin 6 (IL-6), nitric oxide, and total antioxidant capacity. Phytoniosomes were able to protect cells from apoptosis by decreasing caspase 3 activity. Network pharmacology and bioinformatic analysis confirmed the induction of the effect via Akt-PI3K-MAPK and AGE-RAGE signalling pathways through quercetin and luteolin actions. CONCLUSION: The current report highlights the potential of Artemisia phytoniosomes as strong contenders in AGE-related disease therapy.


Subject(s)
Artemisia , Diabetes Mellitus , Drugs, Chinese Herbal , Antioxidants/pharmacology , Artemisia/chemistry , Caspase 3 , Chromatography, Liquid , Interleukin-6 , Liver/metabolism , Luteolin , Network Pharmacology , Nitric Oxide , Phosphatidylinositol 3-Kinases , Plant Extracts/chemistry , Plant Extracts/pharmacology , Proto-Oncogene Proteins c-akt/metabolism , Quercetin , Solvents , Tandem Mass Spectrometry/methods , Tumor Necrosis Factor-alpha
4.
Mol Cell Biochem ; 477(10): 2345-2357, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35543857

ABSTRACT

Artemisia herba-alba (AHA) is a traditionally used plant to treat various diseases, including diabetes and metabolic dysfunctions. Plant extracts are generally explored empirically without a deeper assessment of their mechanism of action. Here, we describe a combinatorial study of biochemical, molecular, and bioinformatic (metabolite-protein pharmacology network) analyses to elucidate the mechanism of action of AHA and shed light on its multilevel effects in the treatment of diabetes-related advanced glycation end-products (AGE)-induced liver damages. The extract's polyphenols and flavonoids content were measured and then identified via LC-Q-TOF-MS/MS. Active compounds were used to generate a metabolite-target interaction network via Swiss Target Prediction and other databases. The extract was tested for its antiglycation and aggregation properties. Next, THLE-2 liver cells were challenged with AGEs, and the mechanistic markers were measured [TNF-α, IL-6, nitric oxide, total antioxidant capacity, lipid peroxidation (LPO), and caspase 3]. Metabolite and network screening showed the involvement of AHA in diabetes, glycation, liver diseases, aging, and apoptosis. Experimental confirmation showed that AHA inhibited protein modification and AGE formation. Additionally, AHA reduced inflammatory mediators (IL-6, TNFα), oxidative stress markers (NO, LPO), and apoptosis (Caspase 3). On the other hand, cellular total antioxidant capacity was restored to normal levels. The combinatorial study showed that AHA regulates AGE-induced liver damages through MAPK-AKT and AGE-RAGE signaling pathways. This report highlights the combination of experimental and network pharmacology for the exact elucidation of AHA mechanism of action as a multitarget option in the therapy of diabetes and AGEs-related diseases.


Subject(s)
Artemisia , Diabetes Mellitus , Antioxidants/pharmacology , Artemisia/metabolism , Caspase 3/metabolism , Diabetes Mellitus/drug therapy , Flavonoids/pharmacology , Glycation End Products, Advanced/metabolism , Inflammation Mediators/metabolism , Interleukin-6/metabolism , Liver/metabolism , Nitric Oxide/metabolism , Plant Extracts/pharmacology , Polyphenols/pharmacology , Proto-Oncogene Proteins c-akt/metabolism , Receptor for Advanced Glycation End Products/metabolism , Signal Transduction , Tandem Mass Spectrometry , Tumor Necrosis Factor-alpha/metabolism
5.
Mikrochim Acta ; 189(5): 202, 2022 04 26.
Article in English | MEDLINE | ID: mdl-35474492

ABSTRACT

The construction of a rapid and easy immunofluorescence bioassay for SARS-CoV-2 detection is described. We report for the first time a novel one-pot synthetic approach for simultaneous photoinduced step-growth polymerization of pyrene (Py) and ring-opening polymerization of ε-caprolactone (PCL) to produce a graft fluorescent copolymer PPy-g-PCL that was conjugated to SARS-CoV-2-specific antibodies using EDC/NHS chemistry. The synthesis steps and conjugation products were fully characterized using standard spectral analysis. Next, the PPy-g-PCL was used for the construction of a dot-blot assay which was calibrated for applications to human nasopharyngeal samples. The analytical features of the proposed sensor showed a detection range of 6.03-8.7 LOG viral copy mL-1 (Ct Scores: 8-25), the limit of detection (LOD), and quantification (LOQ) of 1.84 and 6.16 LOG viral copy mL-1, respectively. The repeatability and reproducibility of the platform had a coefficient of variation (CV) ranging between 1.2 and 5.9%. The fluorescence-based dot-blot assay was tested with human samples. Significant differences were observed between the fluorescence intensity of the negative and positive samples, with an overall correct response of 93.33%. The assay demonstrated a high correlation with RT-PCR data. This strategy opens new insights into simplified synthesis procedures of the reporter molecules and their high potential sensing and diagnosis applications.


Subject(s)
COVID-19 , SARS-CoV-2 , Biological Assay , COVID-19/diagnosis , Caproates , Coloring Agents , Humans , Lactones , Poly A , Polyesters , Polymerization , Reproducibility of Results
6.
Talanta ; 243: 123356, 2022 Jun 01.
Article in English | MEDLINE | ID: mdl-35248943

ABSTRACT

The increasing mutation frequency of the SARS-CoV-2 virus and the emergence of successive variants have made correct diagnosis hard to perform. Developing efficient and accurate methods to diagnose infected patients is crucial to effectively mitigate the pandemic. Here, we developed an electrochemical immunosensor based on SARS-CoV-2 antibody cocktail-conjugated magnetic nanoparticles for the sensitive and accurate detection of the SARS-CoV-2 virus and its variants in nasopharyngeal swabs. The application of the antibody cocktail was compared with commercially available anti-SARS-CoV-2 S1 (anti-S1) and anti-S2 monoclonal antibodies. After optimization and calibration, the limit of detection (LOD) determination demonstrated a LOD = 0.53-0.75 ng/mL for the antibody cocktail-based sensor compared with 0.93 ng/mL and 0.99 ng/mL for the platforms using anti-S1 and anti-S2, respectively. The platforms were tested with human nasopharyngeal swab samples pre-diagnosed with RT-PCR (10 negatives and 40 positive samples). The positive samples include the original, alpha, beta, and delta variants (n = 10, for each). The polyclonal antibody cocktail performed better than commercial anti-S1 and anti-S2 antibodies for all samples reaching 100% overall sensitivity, specificity, and accuracy. It also showed a wide range of variants detection compared to monoclonal antibody-based platforms. The present work proposes a versatile electrochemical biosensor for the indiscriminate detection of the different variants of SARS-CoV-2 using a polyclonal antibody cocktail. Such diagnostic tools allowing the detection of variants can be of great efficiency and economic value in the fight against the ever-changing SARS-CoV-2 virus.


Subject(s)
Biosensing Techniques , COVID-19 , Magnetite Nanoparticles , COVID-19/diagnosis , Humans , Immunoassay , SARS-CoV-2/genetics
7.
Biosens Bioelectron X ; 10: 100105, 2022 May.
Article in English | MEDLINE | ID: mdl-35036904

ABSTRACT

Point of care (PoC) devices are highly demanding to control current pandemic, originated from severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2). Though nucleic acid-based methods such as RT-PCR are widely available, they require sample preparation and long processing time. PoC diagnostic devices provide relatively faster and stable results. However they require further investigation to provide high accuracy and be adaptable for the new variants. In this study, laser-scribed graphene (LSG) sensors are coupled with gold nanoparticles (AuNPs) as stable promising biosensing platforms. Angiotensin Converting Enzyme 2 (ACE2), an enzymatic receptor, is chosen to be the biorecognition unit due to its high binding affinity towards spike proteins as a key-lock model. The sensor was integrated to a homemade and portable potentistat device, wirelessly connected to a smartphone having a customized application for easy operation. LODs of 5.14 and 2.09 ng/mL was achieved for S1 and S2 protein in the linear range of 1.0-200 ng/mL, respectively. Clinical study has been conducted with nasopharyngeal swabs from 63 patients having alpha (B.1.1.7), beta (B.1.351), delta (B.1.617.2) variants, patients without mutation and negative patients. A machine learning model was developed with accuracy of 99.37% for the identification of the SARS-Cov-2 variants under 1 min. With the increasing need for rapid and improved disease diagnosis and monitoring, the PoC platform proved its potential for real time monitoring by providing accurate and fast variant identification without any expertise and pre sample preparation, which is exactly what societies need in this time of pandemic.

8.
Anal Bioanal Chem ; 413(29): 7251-7263, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34622322

ABSTRACT

Supply shortage for the development and production of preventive, therapeutic, and diagnosis tools during the COVID-19 pandemic is an important issue affecting the wealthy and poor nations alike. Antibodies and antigens are especially needed for the production of immunological-based testing tools such as point-of-care tests. Here, we propose a simple and quick magnetic nanoparticle (MNP)-based separation/isolation approach for the repurposing of infected human samples to produce specific antibodies and antigen cocktails. Initially, an antibody cocktail was purified from serums via precipitation and immunoaffinity chromatography. Purified antibodies were conjugated onto MNPs and used as an affinity matrix to separate antigens. The characterization process was performed by ELISA, SDS-PAGE, electrochemistry, isothermal titration calorimetry, and LC-Q-TOF-MS/MS analyses. The MNP-separated peptides can be used for mass spectrometry-based as well as paper-based lateral flow assay diagnostic. The exploitation of the current workflow for the development of efficient diagnostic tools, specific treatments, and fundamental research can significantly impact the present or eventual pandemic. This workflow can be considered as a two birds, one stone-like strategy.


Subject(s)
Antibodies, Viral/isolation & purification , Antigens, Viral/isolation & purification , COVID-19/diagnosis , Cost-Benefit Analysis , Immunoassay/economics , SARS-CoV-2/isolation & purification , Viremia/virology , Antibodies, Viral/blood , Antigens, Viral/blood , COVID-19/virology , Calorimetry , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Humans , SARS-CoV-2/immunology , Specimen Handling , Tandem Mass Spectrometry , Viremia/blood , Workflow
9.
ACS Sens ; 6(8): 2988-2997, 2021 08 27.
Article in English | MEDLINE | ID: mdl-34270230

ABSTRACT

The global pandemic of COVID-19 continues to be an important threat, especially with the fast transmission rate observed after the discovery of novel mutations. In this perspective, prompt diagnosis requires massive economical and human resources to mitigate the disease. The current study proposes a rational design of a colorimetric lateral flow immunoassay (LFA) based on the repurposing of human samples to produce COVID-19-specific antigens and antibodies in combination with a novel dye-loaded polymersome for naked-eye detection. A group of 121 human samples (61 serums and 60 nasal swabs) were obtained and analyzed by RT-PCR and ELISA. Pooled samples were used to purify antibodies using affinity chromatography, while antigens were purified via magnetic nanoparticles-based affinity. The purified proteins were confirmed for their specificity to COVID-19 via commercial LFA, ELISA, and electrochemical tests in addition to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Polymersomes were prepared using methoxy polyethylene glycol-b-polycaprolactone (mPEG-b-PCL) diblock copolymers and loaded with a Coomassie Blue dye. The polymersomes were then functionalized with the purified antibodies and applied for the preparation of two types of LFA (antigen test and antibody test). Overall, the proposed diagnostic tests demonstrated 93 and 92.2% sensitivity for antigen and antibody tests, respectively. The repeatability (92-94%) and reproducibility (96-98%) of the tests highlight the potential of the proposed LFA. The LFA test was also analyzed for stability, and after 4 weeks, 91-97% correct diagnosis was observed. The current LFA platform is a valuable assay that has great economical and analytical potential for widespread applications.


Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Viral , COVID-19 Testing , Humans , Reproducibility of Results
10.
Bioconjug Chem ; 32(8): 1491-1502, 2021 08 18.
Article in English | MEDLINE | ID: mdl-34283580

ABSTRACT

Chemotherapy has seen great progress in the development of performant treatment strategies. Nanovesicles such as liposomes and polymersomes demonstrated great potential in cancer therapy. However, these nanocarriers deliver their content passively, which faces a lot of constraints during blood circulation. The main challenge resides in degradation and random delivery to normal tissues. Hence, targeting drug delivery using specific molecules (such as antibodies) grafted over the surface of these nanocarriers came as the answer to overcome many problems faced before. The advantage of using antibodies is their antigen/antibody recognition, which provides a high level of specificity to reach treatment targets. This review discusses the many techniques of nanocarrier functionalization with antibodies. The aim is to recognize the various approaches by describing their advantages and deficiencies to create the most suitable drug delivery platform. Some methods are more suitable for other applications rather than drug delivery, which can explain the low success of some proposed targeted nanocarriers. In here, a critical analysis of how every method could impact the recognition and targeting capacity of some nanocarriers (liposomes and polymersomes) is discussed to make future research more impactful and advance the field of biomedicine further.


Subject(s)
Drug Delivery Systems , Liposomes/chemistry , Pharmaceutical Preparations/administration & dosage , Animals , Antibodies/chemistry , Drug Delivery Systems/methods , Humans , Nanoparticles/chemistry , Polymers/chemistry , Surface Properties
11.
Biosens Bioelectron ; 192: 113484, 2021 Nov 15.
Article in English | MEDLINE | ID: mdl-34246807

ABSTRACT

Real-time reverse transcriptase-polymerase chain reaction (RT-PCR)-based assays are the gold standard for virus diagnosis. Point-of-care (POC) technologies have shown great progress during this period. Herein, we propose a novel fuchsine dye-loaded polymersome for a colorimetric paper-based dot blot spike protein diagnostic assay for COVID-19 via smartphone-assisted sensing. The prepared platform aimed to create an adaptable tool that competes with traditional nanoparticle-based assays employing gold and silver. Analytical characterization and application of the testing platform showed high sensitivity (10 times better than gold nanoparticles), stability, fast turnaround, and reproducibility. The potential and possibilities demonstrated by the current platform could be observed in its adaptability for different markers and pathologies. In addition, smartphone-assisted sensing emphasizes the ability to use the tool at home by common peoples which can lower the burden on the healthcare facilities and reach more underdeveloped regions.


Subject(s)
Biosensing Techniques , COVID-19 , Spike Glycoprotein, Coronavirus/analysis , COVID-19/diagnosis , Gold , Humans , Metal Nanoparticles , Reproducibility of Results , Rosaniline Dyes , SARS-CoV-2 , Sensitivity and Specificity
12.
Anal Chem ; 93(24): 8585-8594, 2021 06 22.
Article in English | MEDLINE | ID: mdl-34081452

ABSTRACT

The global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has revealed the urgent need for accurate, rapid, and affordable diagnostic tests for epidemic understanding and management by monitoring the population worldwide. Though current diagnostic methods including real-time polymerase chain reaction (RT-PCR) provide sensitive detection of SARS-CoV-2, they require relatively long processing time, equipped laboratory facilities, and highly skilled personnel. Laser-scribed graphene (LSG)-based biosensing platforms have gained enormous attention as miniaturized electrochemical systems, holding an enormous potential as point-of-care (POC) diagnostic tools. We describe here a miniaturized LSG-based electrochemical sensing scheme for coronavirus disease 2019 (COVID-19) diagnosis combined with three-dimensional (3D) gold nanostructures. This electrode was modified with the SARS-CoV-2 spike protein antibody following the proper surface modifications proved by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) characterizations as well as electrochemical techniques. The system was integrated into a handheld POC detection system operated using a custom smartphone application, providing a user-friendly diagnostic platform due to its ease of operation, accessibility, and systematic data management. The analytical features of the electrochemical immunoassay were evaluated using the standard solution of S-protein in the range of 5.0-500 ng/mL with a detection limit of 2.9 ng/mL. A clinical study was carried out on 23 patient blood serum samples with successful COVID-19 diagnosis, compared to the commercial RT-PCR, antibody blood test, and enzyme-linked immunosorbent assay (ELISA) IgG and IgA test results. Our test provides faster results compared to commercial diagnostic tools and offers a promising alternative solution for next-generation POC applications.


Subject(s)
Biosensing Techniques , COVID-19 , Graphite , Point-of-Care Systems , Antibodies, Viral , COVID-19/diagnosis , COVID-19 Testing , Gold , Humans , Lasers , Nanostructures , SARS-CoV-2 , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus
13.
Talanta ; 222: 121534, 2021 Jan 15.
Article in English | MEDLINE | ID: mdl-33167242

ABSTRACT

As COVID-19 has reached pandemic status and the number of cases continues to grow, widespread availability of diagnostic testing is critical in helping identify and control the emergence of this rapidly spreading and serious illness. However, a lacking in making a quick reaction to the threat and starting early development of diagnostic sensing tools has had an important impact globally. In this regard, here we will review critically the current developed diagnostic tools in response to the COVID-19 pandemic and compare the different types through the discussion of their pros and cons such as nucleic acid detection tests (including PCR and CRISPR), antibody and protein-based diagnosis tests. In addition, potential technologies that are under development such as on-site diagnosis platforms, lateral flow, and portable PCR units are discussed. Data collection and epidemiological analysis could also be an interesting factor to incorporate with the emerging technologies especially with the wide access to smartphones. Lastly, a SWOT analysis and perspectives on how the development of novel sensory platforms should be treated by the different decision-makers are analyzed.


Subject(s)
Clinical Laboratory Techniques/methods , Coronavirus Infections/diagnosis , Coronavirus Infections/epidemiology , Pandemics , Pneumonia, Viral/diagnosis , Pneumonia, Viral/epidemiology , COVID-19 , COVID-19 Testing , Clinical Laboratory Techniques/instrumentation , Humans , Point-of-Care Testing , Reagent Kits, Diagnostic
14.
Arch Biochem Biophys ; 695: 108643, 2020 11 30.
Article in English | MEDLINE | ID: mdl-33122162

ABSTRACT

Artificial nanostructures using polymers to produce polymeric vesicles are inspired by the many intricate structures found in living organisms. Polymersomes are a class of self-assembled vesicles known for their great stability and application in drug delivery. They can be tuned according to their intended use by changing their components and introducing activable block copolymers that transform these polymersomes into smart nanocarriers. In this study, we propose the synthesis of a poly (ethylene oxide)-poly (ε-caprolactone)-based polymersome (PEO-PCL) loaded with GSH as a pH-responsive drug delivery molecule for cancer and protein alteration inhibition. Initially, the nanocarrier was synthesized and characterized by DLS, TEM/SEM microscopy as well as gel permeation chromatography (GPC) and 1H NMR. Their CMC formation, encapsulation efficiency, and pH responsiveness were analyzed. In addition, empty and GSH-loaded PEO-PCL polymersomes were tested for their toxicity and therapeutic effect on normal and cancer cells via an MTT test. Subsequently, protein alteration models (aggregation, glycation, and oxidation) were performed in vitro where the polymersomes were tested. Results showed that other than being non-toxic and able to highly encapsulate and release the GSH in response to acidic conditions, the nanocomposites do not hinder its content's ameliorative effects on cancer cells and protein alterations. This infers that polymeric nanocarriers can be a base for future smart biomedicine applications and theranostics.


Subject(s)
Drug Carriers , Glutathione , Neoplasm Proteins/metabolism , Neoplasms , Polyesters , Animals , Chlorocebus aethiops , Delayed-Action Preparations/chemistry , Delayed-Action Preparations/pharmacokinetics , Delayed-Action Preparations/pharmacology , Drug Carriers/chemistry , Drug Carriers/pharmacokinetics , Drug Carriers/pharmacology , Glutathione/chemistry , Glutathione/pharmacokinetics , Glutathione/pharmacology , Glycosylation/drug effects , HeLa Cells , Humans , Hydrogen-Ion Concentration , Neoplasms/drug therapy , Neoplasms/metabolism , Neoplasms/pathology , Polyesters/chemistry , Polyesters/pharmacokinetics , Polyesters/pharmacology , Vero Cells
15.
Biometals ; 33(6): 353-364, 2020 12.
Article in English | MEDLINE | ID: mdl-32997290

ABSTRACT

Age-related complications including protein alterations seen in diabetes and Alzheimer's disease are a major issue due to their accumulation and deleterious effects. This report aims to investigate the effect of zinc supplementation on the anti-glycoxidation activity of carnosine on the in vitro model of albumin-based protein modification. Besides, the therapeutic effect of this combination was tested through the addition of the molecules in tandem (co-treatment) or post initiation (post-treatment) of the protein modification process. Glycation was induced via the addition of glucose to which carnosine (5 mM) alone or with various zinc concentrations (125, 250, and 500 µM) were added either at 0 h or 24 h post-glycation induction. On the other hand, protein oxidation was induced using chloramine T (20 mM) and treated in the same way with carnosine and zinc. The different markers of glycation (advanced glycation end products (AGEs), dityrosine, and beta-sheet formation (aggregation)) and oxidation (AOPP, advanced oxidation protein products) were estimated via fluorescence and colorimetric assays. Zinc addition induced a significant enhancement of carnosine activity by reducing albumin modification that outperformed aminoguanidine both in the co- and post-treatment protocols. Zinc demonstrated a supplementary effect in combination with carnosine highlighting its potential in the protection against age-related protein modifications processes such as the ones found in diabetes.


Subject(s)
Carnosine/pharmacology , Models, Biological , Serum Albumin, Bovine/antagonists & inhibitors , Zinc/pharmacology , Animals , Cattle , Glycosylation , Oxidation-Reduction , Serum Albumin, Bovine/metabolism
16.
Biomater Sci ; 8(12): 3237-3247, 2020 Jun 21.
Article in English | MEDLINE | ID: mdl-32484498

ABSTRACT

Nanotechnology has seen an outburst in biomedicine applications through the use of nanoparticles of different sources in therapy and diagnostics. The needs of theranostics evolved through the years for the development of tailored treatments. In this regard, nanocarriers have shown a great impact on the field via the use of natural lipidic vesicles for drug delivery. This breakthrough allowed the medical field to protect the drug from undesired interactions in the bloodstream and lowered the drug load usually given to reach therapeutical doses. Nanocarriers further continued by using block polymers to create more stable structures with higher protection levels of their content. In this review, we introduce both lipidic and polymeric vesicles with their specific characteristics and discuss the advantages and disadvantages of each type which was taken as a base to introduce the newly known lipid-polymer hybrids that take the advantages from both sides to present an interesting approach to regulate the physicochemical features, pharmacokinetics and other parameters used in tailoring treatments for cancer therapy. In addition, from the many hybrids proposed we have focused our efforts in discussing two major groups that are lipid-polymer hybrid nanoparticles LPHNs (polymersomes inside liposomes), and capsosomes (liposomes inside polymersomes) showing the many potential benefits of combining lipids and polymers for biomedicine.


Subject(s)
Drug Carriers , Nanoparticles , Animals , Drug Carriers/administration & dosage , Drug Carriers/chemistry , Humans , Lipids/administration & dosage , Lipids/chemistry , Nanoparticles/administration & dosage , Nanoparticles/chemistry , Polymers/administration & dosage , Polymers/chemistry
17.
J Food Biochem ; 44(5): e13179, 2020 05.
Article in English | MEDLINE | ID: mdl-32153045

ABSTRACT

Marine organisms represent a great source of natural bioactive molecules, from which bioactive peptides are of great importance in biomedicine application in many diseases such as diabetes and its related complications. In this study, greater weever (Trachinus Draco) myofibrillar proteins were sequentially hydrolyzed and the different RP-HPLC purified fractions were tested for potential inhibitory activities of ACE and DPP4, in addition to metal chelation and antioxidant activities. Four fractions were found to have high levels of activity (with two peptides being multifunctional) and were subsequently sequenced using the de novo sequencing method. The results indicate that the peptides are novel and highly effective for each related activity compared to reference molecules. The current findings suggest these multifunctional peptides as promising therapeutics against oxidative stress, hypertension, and diabetes. PRACTICAL APPLICATIONS: We have described the finding of two multifunctional bioactive peptides from Trachinus Draco (greater weever) myofibrillar proteins having two or more activities. They have ACE inhibitory, DPP4 inhibitory, antioxidant, and metal chelation activities. These new peptides could be used for future biomedicine applications as a stand-alone treatment, in combination with other molecules, or as a supplement. Furthermore, after identification of their sequence in our work, it would have a great potential to be artificially synthesized. The field of food supplements could be explored further.


Subject(s)
Antioxidants , Perciformes , Animals , Antioxidants/pharmacology , Chromatography, Reverse-Phase , Dipeptidyl Peptidase 4 , Peptides
18.
Probiotics Antimicrob Proteins ; 12(1): 48-63, 2020 03.
Article in English | MEDLINE | ID: mdl-31001788

ABSTRACT

Peptides are considered very important due to the diversity expressed through their amino acid sequence, structure variation, large spectrum, and their essential role in biological systems. Antimicrobial peptides (AMPs) emerged as a potent tool in therapy owing to their antimicrobial properties but also their ability to trespass the membranes, specificity, and low toxicity. They comprise a variety of peptides from which specific amino acid-rich peptides are of interest to the current review due to their features in metal interaction and cell penetration. Histidine-rich peptides such as Histatins belong to the metal binding salivary residing peptides with efficient antibacterial, antifungal, and wound-healing activities. Furthermore, their ability to activate in acidic environment attracted the attention to their potential in therapy. The current review covers the current knowledge about AMPs and critically assess the potential of associating with metal ions both structurally and functionally. This review provides interesting hints for the advantages provided by AMPs and metal ions in biomedicine, making use of their direct properties in brain diseases therapy or in the creation of new bio-functionalized nanoparticles for cancer diagnosis and treatment.


Subject(s)
Anti-Infective Agents , Brain Diseases/drug therapy , Cations , Drug Delivery Systems , Nanoparticles/chemistry , Pore Forming Cytotoxic Proteins , Amino Acid Sequence , Anti-Infective Agents/administration & dosage , Anti-Infective Agents/chemistry , Cations/chemistry , Humans , Pore Forming Cytotoxic Proteins/administration & dosage , Pore Forming Cytotoxic Proteins/chemistry
19.
Int J Biol Macromol ; 137: 583-591, 2019 Sep 15.
Article in English | MEDLINE | ID: mdl-31276721

ABSTRACT

Protein modification and alteration are important factors in many age-related diseases such as diabetes and Alzheimer's disease. Modifications like the formation of advanced glycation end-products (AGE) and advanced oxidation protein products (AOPP) can cause harm to the organism and may contribute to protein aggregation and amyloid fibrils formation. Carnosine was used as a potential solution for protein modification complications. Furthermore, some organs like the brain are difficult to reach due to the blood-brain barrier. As such, new nano-engineered formulations were sought to bypass unwanted interactions and degradation. Thus, we propose the encapsulation of carnosine in niosomes as a potential solution. Initially, carnosine niosomes were synthesized and characterized. Then, modifications of bovine serum albumin (glycation, oxidation, and aggregation) were induced in vitro where carnosine and carnosine niosomes were added at different concentrations (2.5, 5, and 10 mM) to the reactions. In addition, biocomputational and docking studies were performed to elucidate the potential interactions. Data showed a dose-dependent inhibition of AGE, AOPP, and aggregation for both carnosine and niosome carnosine. Furthermore, the results suggest that carnosine interacts with specific amino acids implicated in the protein modification process. Carnosine nano-formulation shows promising potential in age-related protein modification and needs further exploration of its mechanisms.


Subject(s)
Carnosine/pharmacology , Serum Albumin, Bovine/chemistry , Serum Albumin, Bovine/metabolism , Animals , Capsules , Carnosine/administration & dosage , Cattle , Drug Compounding , Glycation End Products, Advanced/metabolism , Liposomes , Oxidation-Reduction/drug effects , Protein Aggregates/drug effects
20.
J Food Drug Anal ; 27(1): 22-31, 2019 01.
Article in English | MEDLINE | ID: mdl-30648574

ABSTRACT

Substance use disorders are a widely recognized problem, which affects various levels of communities and influenced the world socioeconomically. Its source is deeply embedded in the global population. In order to fight against such an adversary, governments have spared no efforts in implementing substance abuse treatment centers and funding research to develop treatments and prevention procedures. In this review, we will discuss the use of immunological-based treatments and detection kit technologies. We will be detailing the steps followed to produce performant antibodies (antigens, carriers, and adjuvants) focusing on cocaine and methamphetamine as examples. Furthermore, part of this review is dedicated to substance use detection. Owing to novel technologies such as bio-functional polymeric surfaces and biosensors manufacturing, detection has become a more convenient method with the fast and on-site developed devices. Commercially available devices are able to test substance use disorders in urine, saliva, hair, and sweat. This improvement has had a tremendous impact on the prevention of driving under influence and other illicit behaviors. Lastly, substance abuse became a major issue involving the cooperation of experts on all levels to devise better treatment programs and prevent abuse-based accidents, injury and death.


Subject(s)
Illicit Drugs/analysis , Immunologic Techniques/methods , Substance-Related Disorders/diagnosis , Substance-Related Disorders/therapy , Animals , Humans , Illicit Drugs/blood , Illicit Drugs/urine , Immunologic Techniques/trends , Saliva/chemistry , Substance-Related Disorders/prevention & control , Sweat/chemistry
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