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1.
Sci Rep ; 14(1): 17160, 2024 Jul 26.
Article in English | MEDLINE | ID: mdl-39060395

ABSTRACT

Wireless sensor networks' most prominent concern is energy optimization. It faces significant problems like high energy consumption, data loss, delay, and low network lifetime. To improve, it uses clustering. However, during clustering, coverage holes are most likely to appear near the network's edge, within the cluster, and between clusters. As a result, there are more energy holes and dead nodes; therefore, the goal of this work is to maximize node network lifetime and minimize energy consumption during data transmission in the wireless sensor network (WSN). The proposed work includes three entities: sensor nodes, an edge-assisted unmanned aerial vehicle (UAV), and a base station. It uses an edge-assisted unmanned aerial vehicle to provide additional resources to the UAV, which helps reduce energy consumption during data transmission. This research proposes using communication to enhance the speed and bandwidth of data transmission and reduce transmission latency. This work attempts to improve performance by increasing throughput.

2.
Sci Rep ; 14(1): 12460, 2024 May 30.
Article in English | MEDLINE | ID: mdl-38816518

ABSTRACT

The Schockley-Quisser (SQ) limit of 28.64% is distant from the Sb2S3 solar cells' record power conversion efficiency (PCE), which is 8.00%. Such poor efficiency is mostly owing to substantial interface-induced recombination losses caused by defects at the interfaces and misaligned energy levels. The endeavor of this study is to investigate an efficient Sb2S3 solar cell structure via accurate analytical modeling. The proposed model considers different recombination mechanisms such as non-radiative recombination, Sb2S3/CdS interface recombination, Auger, SRH, tunneling-enhanced recombination, and their combined impact on solar cell performance. This model is verified against experimental work (Glass/ITO/CdS/Sb2S3/Au) where a good coincidence is achieved. Several parameters effects such as thickness, doping, electronic affinity, and bandgap are scrutinized. The effect of both bulk traps located in CdS and Sb2S3 on the electrical outputs of the solar cell is analyzed thoroughly. Besides, a deep insight into the effect of interfacial traps on solar cell figures of merits is gained through shedding light into their relation with carriers' minority lifetime, diffusion length, and surface recombination velocity. Our research findings illuminate that the primary contributors to Sb2S3 degradation are interfacial traps and series resistance. Furthermore, achieving optimal band alignment by fine-tuning the electron affinity of CdS to create a Spike-like conformation is crucial for enhancing the immunity of the device versus the interfacial traps. In our study, the optimized solar cell configuration (Glass/ITO/CdS/Sb2S3/Au) demonstrates remarkable performance, including a high short-circuit current (JSC) of 47.9 mA/cm2, an open-circuit voltage (VOC) of 1.16 V, a fill factor (FF) of 54%, and a notable improvement in conversion efficiency by approximately 30% compared to conventional solar cells. Beyond its superior performance, the optimized Sb2S3 solar cell also exhibits enhanced reliability in mitigating interfacial traps at the CdS/Sb2S3 junction. This improved reliability can be attributed to our precise control of band alignment and the fine-tuning of influencing parameters.

3.
Vet Ital ; 58(2)2022 Dec 30.
Article in English | MEDLINE | ID: mdl-36586114

ABSTRACT

The orf virus (ORFV) is an epitheliotropic virus causing a highly contagious skin disease mainly in sheep and goats. Several diagnostics including molecular tools like Loop mediated isothermal amplification (LAMP) assay are available to detect ORFV in affected species. However, the carry-over contamination associated with LAMP as open tube format prevents the assay applicability as point of care test in field diagnostic settings. In this study, the B2L gene based LAMP assay was optimized in a closed tube format using hydroxynaphthol blue (HNB) and calcein as pre-addition dyes and it has shown a clear positive and negative signal at 60 °C using 4 and 5 mM concentrations of MgSO4 respectively for these dyes. Optimitimzed assay that could reveal the result within one hour is highly specific and senstive with a limit of detection at 12.5 femtogram of viral genomic DNA or ~85 virus genome equivalent. This improved method prevented the cross-contamination of future LAMP reactions in the laboratory without compromising diagnostic sensitivity (100%) and specificity (100%) when compared to open tube system. This closed tube LAMP method has potential to act as a simple visual detection assay for the rapid and specific diagnosis of ORFV in sheep and goats.


Subject(s)
Orf virus , Animals , Sheep , Orf virus/genetics , Goats , Sensitivity and Specificity , Nucleic Acid Amplification Techniques/veterinary , Nucleic Acid Amplification Techniques/methods , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/veterinary , Coloring Agents
4.
Nat Prod Res ; 35(18): 3156-3160, 2021 Sep.
Article in English | MEDLINE | ID: mdl-31711318

ABSTRACT

Phytochemical investigation of the stem bark of Hopea parviflora resulted in the isolation of 9 compounds; which includes friedelin (1), friedelin-3ß-ol (2), (-)-ampelopsin A (3), (-)-ɛ-viniferin (4), (-)-hopeaphenol (5), vaticaphenol A (6), 2,4,8-trihydroxyphenanthrene-2-O-glucoside (7), ellagic acid-3,3',4-trimethoxy-4'-O-α-L-rhamnopyranoside (8) and ß-sitosterol-ß-D-glucoside (9). Among them, compounds 1, 2, 6, 7, 8 and 9 are isolated for the first time from this species. Further, we evaluated the anti-inflammatory activity of compounds 4, 5, 6, 7 and 8. In this study, compound 8 inhibited the activity of proinflammatory mediators like NO, TNF-α, IL-6, 5-LOX and COX-2, also promoted the action of anti-inflammatory mediator like IL-10 via inhibition of the NF-κB pathway in LPS-stimulated RAW 264.7 macrophages.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Dipterocarpaceae , Ellagic Acid/pharmacology , Animals , Anti-Inflammatory Agents/isolation & purification , Dipterocarpaceae/chemistry , Ellagic Acid/isolation & purification , Lipopolysaccharides , Macrophages/drug effects , Mice , NF-kappa B , Phytochemicals/isolation & purification , Phytochemicals/pharmacology , Plant Bark/chemistry , RAW 264.7 Cells
5.
Contemp Clin Dent ; 11(1): 3-9, 2020.
Article in English | MEDLINE | ID: mdl-33110301

ABSTRACT

The association of genetic polymorphisms with periodontitis has been studied extensively. The interleukin-7 (IL-17) is a group of cytokines, which comprises six different molecules (IL-17A, B, C, D, E, and F). Among this, IL-17A is the most commonly understood cytokine, and its polymorphism plays a critical role in inflammatory diseases and periodontal inflammation. The present study was aimed at pooling the data available for meta-analysis and to evaluate whether IL-17A (rs2275913) polymorphism is associated with the susceptibility of chronic periodontitis.

6.
Org Biomol Chem ; 17(21): 5212-5222, 2019 05 29.
Article in English | MEDLINE | ID: mdl-31080990

ABSTRACT

The carbonyl moiety is one of the indispensable sub-units in organic synthesis with significant applications in medicinal as well as materials chemistry. Hence the insertion of a carbonyl group via simple and highly efficient routes has been one of the most challenging tasks for organic chemists. Though the direct utilisation of CO gas in carbonylation is the fundamental procedure for the construction of carbonyl compounds, it has certain drawbacks due to its toxic and explosive nature. As a result, the need for cheap and efficient CO surrogates has gained much attention nowadays by which CO gas can be easily generated in situ or ex situ. In this review we discuss the advantages of chloroform as CO surrogate and have surveyed recent carbonylation reactions where chloroform has been used as CO source.

7.
J Ethnopharmacol ; 236: 196-204, 2019 May 23.
Article in English | MEDLINE | ID: mdl-30844488

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Hopea ponga (Dennst.) Mabb. Is used in traditional herbal formulations for diabetes complications. The aim of this study is to evaluate the antidiabetic effect of extracts and compounds from H. ponga. MATERIALS AND METHODS: Silica gel column chromatography was performed to identify various chemical components of the plant extract. Different extracts of H. ponga and isolated compounds were screened for their antidiabetic effect by modulation of digestive enzymes and protein glycation. The effect of glucose uptake by the compounds and the pathways through which the compounds mediate the glucose uptake potential were confirmed by fluorescent microscopy, flow cytometry and western blot analysis. RESULTS: Acetone and ethanol extracts of the stem bark of Hopea ponga (Dennst.) Mabb. Afforded six resveratrol oligomers namely, E-resveratrol (1), (-)-ε-viniferin (2), (-)-α-viniferin (3), trihydroxyphenanthrene glucoside (THPG) (4), vaticaphenol A (5), (-)-hopeaphenol (6), along with four phytosterols. The structures were determined on the basis of spectroscopic analyses including nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry (HRMS) data. Compounds 1-5 and 7-10 were tested for their α-glucosidase, α-amylase and glycation inhibitiory activities. All the resveratrol oligomers (1-5) showed prominent α-glucosidase inhibition with IC50 values, 12.56 ±â€¯1.00, 23.98 ±â€¯1.11, 7.17 ±â€¯1.10, 31.74 ±â€¯0.42 and 16.95 ±â€¯0.39 µM, respectively. Molecular docking studies also supported the observed α-glucosidase inhibition. Compound 3 displayed IC50 values of 4.85 ±â€¯0.06 and 27.10 ±â€¯0.04 µM in α-amylase and glycation inhibitory assays activity. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay revealed that the compounds 3 and 4 were found to be less toxic at a concentration of 100 µM (<10%) and 25 µM (<20%), respectively. The effect of glucose uptake performed by 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) in L6 myoblast were measured by fluorescent microscopy and flow cytometry. The compounds 3 and 4 showed 2-NBDG uptake of 49.6% and 38.8% respectively. By examining the molecular pathway through which the compounds elicit their glucose uptake potential, it was observed that both the compounds mainly act via AMPK pathway. CONCLUSION: This is the first report on the isolation of compounds from H. ponga. Altogether, the results of this study reveal the antidiabetic effects of H. ponga extracts and isolated compounds promoting traditional use of this plant in the treatment of diabetes.


Subject(s)
Dipterocarpaceae/chemistry , Hypoglycemic Agents/pharmacology , Plant Extracts/pharmacology , Resveratrol/pharmacology , Acetone/chemistry , Animals , Cell Line , Diabetes Mellitus/drug therapy , Enzyme Assays , Ethanol/chemistry , Glycosylation/drug effects , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/isolation & purification , Inhibitory Concentration 50 , Medicine, Traditional/methods , Molecular Docking Simulation , Molecular Structure , Myoblasts , Plant Bark/chemistry , Plant Extracts/chemistry , Plant Stems/chemistry , Rats , Resveratrol/chemistry , Resveratrol/isolation & purification , Toxicity Tests , alpha-Amylases/antagonists & inhibitors , alpha-Amylases/chemistry , alpha-Amylases/metabolism , alpha-Glucosidases/chemistry , alpha-Glucosidases/metabolism
8.
J Thromb Haemost ; 16(5): 946-959, 2018 05.
Article in English | MEDLINE | ID: mdl-29512284

ABSTRACT

Essentials Heat shock protein 47 (HSP47), a collagen specific chaperone is present on the platelet surface. Collagen mediated platelet function was reduced following blockade or deletion of HSP47. GPVI receptor regulated signalling was reduced in HSP47 deficient platelets. Platelet HSP47 tethers to exposed collagen thus modulating thrombosis and hemostasis. SUMMARY: Objective Heat shock protein 47 (HSP47) is an intracellular chaperone protein that is vital for collagen biosynthesis in collagen secreting cells. This protein has also been shown to be present on the surface of platelets. Given the importance of collagen and its interactions with platelets in triggering hemostasis and thrombosis, in this study we sought to characterize the role of HSP47 in these cells. Methods and Results The deletion of HSP47 in mouse platelets or its inhibition in human platelets reduced their function in response to collagen and the GPVI agonist (CRP-XL), but responses to thrombin were unaltered. In the absence of functional HSP47, the interaction of collagen with platelets was reduced, and this was associated with reduced GPVI-collagen binding, signalling and platelet activation. Thrombus formation on collagen, under arterial flow conditions, was also decreased following the inhibition or deletion of HSP47, in the presence or absence of eptifibatide, consistent with a role for HSP47 in enhancing platelet adhesion to collagen. Platelet adhesion under flow to von Willebrand factor was unaltered following HSP47 inhibition. Laser-induced thrombosis in cremaster muscle arterioles was reduced and bleeding time was prolonged in HSP47-deficient mice or following inhibition of HSP47. Conclusions Our study demonstrates the presence of HSP47 on the platelet surface, where it interacts with collagen, stabilizes platelet adhesion and increases collagen-mediated signalling and therefore thrombus formation and hemostasis.


Subject(s)
Blood Platelets/metabolism , Carrier Proteins/blood , Collagen/blood , HSP70 Heat-Shock Proteins/blood , Hemostasis , Platelet Activation , Thrombosis/blood , Animals , Blood Platelets/drug effects , Calcium Signaling , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/genetics , Disease Models, Animal , HSP70 Heat-Shock Proteins/antagonists & inhibitors , HSP70 Heat-Shock Proteins/deficiency , HSP70 Heat-Shock Proteins/genetics , Hemostasis/drug effects , Humans , Mice, Inbred C57BL , Mice, Knockout , Mitochondrial Proteins , Platelet Activation/drug effects , Platelet Adhesiveness , Platelet Aggregation Inhibitors/pharmacology , Platelet Membrane Glycoproteins/metabolism , Protein Binding , Thrombosis/genetics , Thrombosis/prevention & control
9.
J Thromb Haemost ; 16(2): 367-377, 2018 02.
Article in English | MEDLINE | ID: mdl-29052936

ABSTRACT

Essentials ERp72 is a thiol isomerase enzyme. ERp72 levels increase at the platelet surface during platelet activation. We generated a humanized monoclonal antibody which blocks ERp72 enzyme activity (anti-ERp72). Anti-ERp72 inhibits platelet functional responses and thrombosis. SUMMARY: Background Within the endoplasmic reticulum, thiol isomerase enzymes modulate the formation and rearrangement of disulfide bonds in newly folded proteins entering the secretory pathway to ensure correct protein folding. In addition to their intracellular importance, thiol isomerases have been recently identified to be present on the surface of a number of cell types where they are important for cell function. Several thiol isomerases are known to be present on the resting platelet surface, including PDI, ERp5 and ERp57, and levels are increased following platelet activation. Inhibition of the catalytic activity of these enzymes results in diminished platelet function and thrombosis. Aim We previously determined that ERp72 is present at the resting platelet surface and levels increase upon platelet activation; however, its functional role on the cell surface was unclear. We aimed to investigate the role of ERp72 in platelet function and its role in thrombosis. Methods Using HuCAL technology, fully humanized Fc-null anti-ERp72 antibodies were generated. Eleven antibodies were screened for their ability to inhibit ERp72 activity and the most potent inhibitory antibody (anti-ERp72) selected for further testing in platelet functional assays. Results and conclusions Anti-ERp72 inhibited platelet aggregation, granule secretion, calcium mobilisation and integrin activation, revealing an important role for extracellular ERp72 in the regulation of platelet activation. Consistent with this, infusion of anti-ERp72 into mice protected against thrombosis.


Subject(s)
Antibodies, Monoclonal, Humanized/pharmacology , Blood Platelets/drug effects , Fibrinolytic Agents/pharmacology , Membrane Glycoproteins/antagonists & inhibitors , Platelet Activation/drug effects , Platelet Aggregation Inhibitors/pharmacology , Protein Disulfide-Isomerases/antagonists & inhibitors , Thrombosis/prevention & control , Animals , Blood Platelets/enzymology , Blood Platelets/immunology , Calcium/blood , Disease Models, Animal , Fibrinogen/metabolism , Humans , Male , Membrane Glycoproteins/blood , Membrane Glycoproteins/immunology , Mice, Inbred C57BL , Platelet Adhesiveness/drug effects , Platelet Aggregation/drug effects , Platelet Glycoprotein GPIIb-IIIa Complex/metabolism , Protein Disulfide-Isomerases/blood , Protein Disulfide-Isomerases/immunology , Signal Transduction/drug effects , Thrombosis/blood , Thrombosis/enzymology
10.
J Thromb Haemost ; 12(8): 1342-52, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24888521

ABSTRACT

BACKGROUND: Integrin-linked kinase (ILK) and its associated complex of proteins are involved in many cellular activation processes, including cell adhesion and integrin signaling. We have previously demonstrated that mice with induced platelet ILK deficiency show reduced platelet activation and aggregation, but only a minor bleeding defect. Here, we explore this apparent disparity between the cellular and hemostatic phenotypes. METHODS: The impact of ILK inhibition on integrin αII b ß3 activation and degranulation was assessed with the ILK-specific inhibitor QLT0267, and a conditional ILK-deficient mouse model was used to assess the impact of ILK deficiency on in vivo platelet aggregation and thrombus formation. RESULTS: Inhibition of ILK reduced the rate of both fibrinogen binding and α-granule secretion, but was accompanied by only a moderate reduction in the maximum extent of platelet activation or aggregation in vitro. The reduction in the rate of fibrinogen binding occurred prior to degranulation or translocation of αII b ß3 to the platelet surface. The change in the rate of platelet activation in the absence of functional ILK led to a reduction in platelet aggregation in vivo, but did not change the size of thrombi formed following laser injury of the cremaster arteriole wall in ILK-deficient mice. It did, however, result in a marked decrease in the stability of thrombi formed in ILK-deficient mice. CONCLUSION: Taken together, the findings of this study indicate that, although ILK is not essential for platelet activation, it plays a critical role in facilitating rapid platelet activation, which is essential for stable thrombus formation.


Subject(s)
Platelet Activation , Protein Serine-Threonine Kinases/metabolism , Thrombosis/enzymology , Animals , Flow Cytometry , Mice , Mice, Transgenic , Platelet Glycoprotein GPIIb-IIIa Complex/metabolism
11.
J Appl Microbiol ; 115(3): 880-7, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23734819

ABSTRACT

AIM: The aim of the present study is to constitutively express heterologous oxalate decarboxylase (OxdC) in Lactobacillus plantarum and to examine its ability to degrade oxalate in vitro for their future therapy against enteric hyperoxaluria. METHOD AND RESULTS: In this study, we generated a recombinant strain of Lb. plantarum to constitutively overexpress B. subtilis oxalate decarboxylase (oxdC) using a host lactate dehydrogenase promoter (PldhL ). The recombinant Lb. plantarum was able to degrade more than 90% oxalate compared to 15% by the wild type. In addition, the recombinant strain also had higher tolerance up to 500 mmol l(-1) oxalate. CONCLUSION: We developed a recombinant Lb. plantarum NC8 that constitutively expressed heterologous oxalate decarboxylase and degraded oxalate efficiently under in vitro conditions. SIGNIFICANCE AND IMPACT OF STUDY: The long-term aim is to develop an efficient strain for future therapy against oxalosis.


Subject(s)
Carboxy-Lyases/metabolism , Lactobacillus plantarum/genetics , Oxalates/metabolism , Carboxy-Lyases/genetics , Lactobacillus plantarum/growth & development , Lactobacillus plantarum/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism
12.
J Thromb Haemost ; 10(2): 278-88, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22168334

ABSTRACT

BACKGROUND: Thiol isomerases are a family of endoplasmic reticulum enzymes which orchestrate redox-based modifications of protein disulphide bonds. Previous studies have identified important roles for the thiol isomerases PDI and ERp5 in the regulation of normal platelet function. AIM: Recently, we demonstrated the presence of a further five thiol isomerases at the platelet surface. In this report we aim to report the role of one of these enzymes - ERp57 in the regulation of platelet function. METHODS/RESULTS: Using enzyme activity function blocking antibodies, we demonstrate a role for ERp57 in platelet aggregation, dense granule secretion, fibrinogen binding, calcium mobilisation and thrombus formation under arterial conditions. In addition to the effects of ERp57 on isolated platelets, we observe the presence of ERp57 in the developing thrombus in vivo. Furthermore the inhibition of ERp57 function was found to reduce laser-injury induced arterial thrombus formation in a murine model of thrombosis. CONCLUSIONS: These data suggest that ERp57 is important for normal platelet function and opens up the possibility that the regulation of platelet function by a range of cell surface thiol isomerases may represent a broad paradigm for the regulation of haemostasis and thrombosis.


Subject(s)
Blood Platelets/enzymology , Protein Disulfide-Isomerases/metabolism , Thrombosis/enzymology , Adenosine Triphosphate/metabolism , Animals , Antibodies/pharmacology , Blood Platelets/drug effects , Calcium/metabolism , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Fibrinogen/metabolism , Humans , Mice , Mice, Inbred C57BL , Platelet Aggregation , Protein Disulfide-Isomerases/antagonists & inhibitors , Protein Disulfide-Isomerases/immunology , Secretory Vesicles/metabolism , Thrombosis/pathology , Thrombosis/prevention & control , Time Factors
13.
Asian Pac J Trop Biomed ; 2(4): 281-6, 2012 Apr.
Article in English | MEDLINE | ID: mdl-23569914

ABSTRACT

OBJECTIVE: To investigate the antidiabetic property of Merremia emarginata (M. emarginata) Burm. F. plant in streptozotocin induced diabetic rats. METHODS: The dose dependent effects of 28 days oral treatment with methanol extract (100, 200 and 400 mg/kg) from the plant of M. emarginata on blood glucose level, body weight, insulin, total hemoglobin, glycosylated haemoglobin (HbA1C), total protein, serum urea, serum creatinine and carbohydrate metabolizing enzymes were evaluated in streptozotocin induced diabetic rats. Histology of pancreas was also studied. RESULTS: A significant decrease in blood glucose, serum urea and serum creatinine and significant increase in body weight, insulin and protein level were observed in diabetic rats treated with M. emarginata. Treatment with M. emarginata resulted in a significant reduction of HbA1C and an increase in total hemoglobin level. The activities of carbohydrate metabolizing enzymes such as hexokinase were significantly increased whereas glucose-6-phosphatase, fructose-1, 6-bisphosphatase were significantly decreased by the administration of M. emarginata in diabetic rats. Histology of diabetic rats treated with M. emarginata showed the pancreatic ß-cells regeneration. CONCLUSIONS: These findings suggest that M. emarginata has potent antidiabetic activity in streptozotocin induced diabetic rats.


Subject(s)
Convolvulaceae/chemistry , Diabetes Mellitus, Experimental/metabolism , Hypoglycemic Agents/pharmacology , Plant Extracts/pharmacology , Animals , Blood Glucose/drug effects , Body Weight/drug effects , Hypoglycemic Agents/chemistry , Insulin/blood , Male , Plant Extracts/chemistry , Rats , Rats, Wistar
14.
Protein Pept Lett ; 10(5): 427-33, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14561130

ABSTRACT

Retro-ACP (74-65) fragment was synthesized on a novel 4% tri-(propylene glycol) glycerolate diacrylate cross-linked polystyrene (PS-TRPGGDA) support. The peptide is grown from the functional site present in the cross-linker, which makes it unique and cost-effective among other styrene based polymer supports. A comparative study with Merrifield resin indicates high yield and purity of the peptide synthesized on the novel support.


Subject(s)
Acyl Carrier Protein/chemical synthesis , Peptide Fragments/chemical synthesis , Polystyrenes/chemical synthesis , Acrylates/chemistry , Amino Acid Sequence , Chromatography, High Pressure Liquid , Cross-Linking Reagents/chemistry , Glycerol/chemistry , Solvents
15.
J Pept Res ; 62(1): 1-10, 2003 Jul.
Article in English | MEDLINE | ID: mdl-12787445

ABSTRACT

A highly flexible, mechanically and chemically stable copolymer, tri(propylene glycol) glycerolate diacrylate cross-linked polystyrene (PS-TRPGGDA), was synthesized by the suspension polymerization and employed as a solid support for peptide synthesis. The beaded polymer support containing secondary hydroxyl functional groups in the cross-linker was used as the growth site for peptide synthesis. The procedure is unique and cost-effective in that it avoids the initial functionalization steps required for most of the styrene-based polymer supports. The resin was characterized by 13C-CP-MAS NMR spectroscopy and the morphologic features of the resin were investigated using scanning electron microscopy. Swelling studies conducted on the new support revealed that the PS-TRPGGDA resin undergoes more effective swelling and solvation than PS-DVB resin in all solvents used in peptide synthesis. The efficiency of the new support was demonstrated by synthesizing a 'difficult' sequence Ala-Arg-(Ala)6-Lys and comparing it with commercially available Merrifield and Sheppard resins. The synthetic efficiency was further demonstrated by the synthesis of a 24-residue NR 2A peptide substrate of calcium/calmodulin-binding peptide. The high yield and purity of the peptide synthesized on the novel support indicates the positive role of the flexible and hydrophilic cross-linking agent in the solid support.


Subject(s)
Acrylates/chemical synthesis , Peptide Biosynthesis , Polystyrenes/chemical synthesis , Resins, Synthetic/chemical synthesis , Chromatography, High Pressure Liquid , Cross-Linking Reagents , Glycerol/chemistry , Magnetic Resonance Spectroscopy , Microscopy, Electron, Scanning , Oligopeptides/chemical synthesis , Propylene Glycol/chemistry , Solvents , Spectroscopy, Fourier Transform Infrared
16.
J Pept Res ; 57(2): 140-50, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11168897

ABSTRACT

Peptides were synthesized with very high purity and yield on a highly solvating copolymer of 1,4-butanediol dimethacrylate cross-linked polystyrene support (PS-BDODMA). The polymer was synthesized by radical aqueous suspension polymerization using toluene as the diluent and 1% polyvinyl alcohol as the suspension stabilizer. The supports were highly resistant to various chemical reagents and solvents that are frequently used in polypeptide synthesis. The peptides synthesized on this support were designed from the core (C), envelope (E1 and E2) and nonstructural protein (NS1-NS5) regions of the prototype hepatitis C viral (HCV) polyprotein, and were used to develop a peptide-based immunoassay (PBEIA) for the detection of HCV antibodies. The purity of these peptides was tested by HPLC. Peptide identity was confirmed by amino acid analysis and MALDI-TOF-MS. A single peptide chosen from a conserved area (E2/NS1) at the C-terminus of the hypervariable region (HVRI) was found to be sufficient for effective and reliable diagnosis of HCV infection in infected individuals, as well as also apparently healthy individuals. The CD spectrum of the peptide shows no preference for any ordered secondary structure. When used, peptide mixtures from various protein regions of HCV reduced the sensitivity and reliability of the diagnosis partly because of epitope masking.


Subject(s)
Conserved Sequence , Hepacivirus/chemistry , Hepatitis C Antibodies/analysis , Immunodominant Epitopes/chemistry , Polystyrenes/chemistry , Amino Acid Sequence , Hepatitis C Antibodies/immunology , Immunodominant Epitopes/immunology , Immunoenzyme Techniques , Molecular Sequence Data
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