In Vitro Anti-Protozoal Activity of Propolis Extract and Cysteine Proteases Inhibitor ( Phenyl Vinyl Sulfone ) on Blastocystis Species

2016 ◽  
Vol 46 (2) ◽  
pp. 261-272 ◽  
Author(s):  
Amira B. Mokhtar ◽  
Eman K. El-Gayar ◽  
Eman S. Habib
Keyword(s):  
Cysteine Proteases ◽  
Vinyl Sulfone ◽  
Propolis Extract ◽  
Phenyl Vinyl

Fasciola gigantica: Evaluation of the effect of phenyl vinyl sulfone in vitro

2008 ◽  
Vol 119 (1) ◽  
pp. 125-134 ◽  
Author(s):  
Moshira M.F. Helmy ◽  
Zeinab H. Fahmy ◽  
Hoda Y. Sabry
Keyword(s):  
Fasciola Gigantica ◽  
Vinyl Sulfone ◽  
Phenyl Vinyl

scholarly journals Antifungal activity of dendritic cell lysosomal proteins against Cryptococcus neoformans

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Benjamin N. Nelson ◽  
Savannah G. Beakley ◽  
Sierra Posey ◽  
Brittney Conn ◽  
Emma Maritz ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Cryptococcus Neoformans ◽  
Mammalian Cells ◽  
Cysteine Proteases ◽  
Dependent Manner ◽  
Life Threatening ◽  
Opportunistic Fungal Pathogen ◽  
Dose Dependent ◽  
Lysosomal Proteins

AbstractCryptococcal meningitis is a life-threatening disease among immune compromised individuals that is caused by the opportunistic fungal pathogen Cryptococcus neoformans. Previous studies have shown that the fungus is phagocytosed by dendritic cells (DCs) and trafficked to the lysosome where it is killed by both oxidative and non-oxidative mechanisms. While certain molecules from the lysosome are known to kill or inhibit the growth of C. neoformans, the lysosome is an organelle containing many different proteins and enzymes that are designed to degrade phagocytosed material. We hypothesized that multiple lysosomal components, including cysteine proteases and antimicrobial peptides, could inhibit the growth of C. neoformans. Our study identified the contents of the DC lysosome and examined the anti-cryptococcal properties of different proteins found within the lysosome. Results showed several DC lysosomal proteins affected the growth of C. neoformans in vitro. The proteins that killed or inhibited the fungus did so in a dose-dependent manner. Furthermore, the concentration of protein needed for cryptococcal inhibition was found to be non-cytotoxic to mammalian cells. These data show that many DC lysosomal proteins have antifungal activity and have potential as immune-based therapeutics.

scholarly journals A novel LC-MS method development and validation for the determination of phenyl vinyl sulfone in eletriptan hydrobromide

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Indhu Priya Mabbu ◽  
G. Sumathi ◽  
N. Devanna
Keyword(s):  
Method Development ◽  
Limit Of Detection ◽  
Vinyl Sulfone ◽  
Limit Of Quantification ◽  
Relative Standard ◽  
Pressure Chemical Ionization ◽  
Pressure Chemical ◽  
Development And Validation ◽  
Phenyl Vinyl

Abstract Background The aim of the present method is to develop and validate a specific, sensitive, precise, and accurate liquid chromatography-mass spectrometry (LC-MS) method for the estimation of the phenyl vinyl sulfone in the eletriptan hydrobromide. The effective separation of the phenyl vinyl sulfone was achieved by the Symmetry C18 (50 × 4.6 mm, 3.5 μm) column and a mobile phase composition of 0.1%v/v ammonia buffer to methanol (5:95 v/v), using 0.45 ml/min flow rate and 20 μl of injection volume, with methanol used as diluent. The phenyl vinyl sulfone was monitored on atomic pressure chemical ionization mode mass spectrometer with positive polarity mode. Results The retention time of phenyl vinyl sulfone was found at 2.13 min. The limit of detection (LOD) and limit of quantification (LOQ) were observed at 1.43 ppm and 4.77 ppm concentration respectively; the linear range was found in the concentration ranges from 4.77 to 27.00 ppm with regression coefficient of 0.9990 and accuracy in the range of 97.50–102.10%. The percentage relative standard deviation (% RSD) for six replicates said to be injections were less than 10%. Conclusion The proposed method was validated successfully as per ICH guidelines. Hence, this is employed for the determination of phenyl vinyl sulfone in the eletriptan hydrobromide.

scholarly journals Reversed Proteolysis—Proteases as Peptide Ligases

Catalysts ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 33
Author(s):  
Peter Goettig
Keyword(s):  
Serine Proteases ◽  
Academic Research ◽  
Building Blocks ◽  
Peptide Bond ◽  
Cysteine Proteases ◽  
Small Peptides ◽  
Natural Peptide ◽  
Peptide Esters ◽  
Natural Amino Acids

Historically, ligase activity by proteases was theoretically derived due to their catalyst nature, and it was experimentally observed as early as around 1900. Initially, the digestive proteases, such as pepsin, chymotrypsin, and trypsin were employed to perform in vitro syntheses of small peptides. Protease-catalyzed ligation is more efficient than peptide bond hydrolysis in organic solvents, representing control of the thermodynamic equilibrium. Peptide esters readily form acyl intermediates with serine and cysteine proteases, followed by peptide bond synthesis at the N-terminus of another residue. This type of reaction is under kinetic control, favoring aminolysis over hydrolysis. Although only a few natural peptide ligases are known, such as ubiquitin ligases, sortases, and legumains, the principle of proteases as general catalysts could be adapted to engineer some proteases accordingly. In particular, the serine proteases subtilisin and trypsin were converted to efficient ligases, which are known as subtiligase and trypsiligase. Together with sortases and legumains, they turned out to be very useful in linking peptides and proteins with a great variety of molecules, including biomarkers, sugars or building blocks with non-natural amino acids. Thus, these engineered enzymes are a promising branch for academic research and for pharmaceutical progress.

scholarly journals Identification of Vinyl Sulfone Derivatives as EGFR Tyrosine Kinase Inhibitor: In Vitro and In Silico Studies

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2211
Author(s):  
Thitinan Aiebchun ◽  
Panupong Mahalapbutr ◽  
Atima Auepattanapong ◽  
Onnicha Khaikate ◽  
Supaphorn Seetaha ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Inhibitory Activity ◽  
In Silico ◽  
Kinase Inhibitor ◽  
Kinase Assay ◽  
Vinyl Sulfone ◽  
Egfr Tyrosine Kinase ◽  
In Silico Studies ◽  
Sulfone Derivatives

Epidermal growth factor receptor (EGFR), overexpressed in many types of cancer, has been proved as a high potential target for targeted cancer therapy due to its role in regulating proliferation and survival of cancer cells. In the present study, a series of designed vinyl sulfone derivatives was screened against EGFR tyrosine kinase (EGFR-TK) using in silico and in vitro studies. The molecular docking results suggested that, among 78 vinyl sulfones, there were eight compounds that could interact well with the EGFR-TK at the ATP-binding site. Afterwards, these screened compounds were tested for the inhibitory activity towards EGFR-TK using ADP-Glo™ kinase assay, and we found that only VF16 compound exhibited promising inhibitory activity against EGFR-TK with the IC50 value of 7.85 ± 0.88 nM. In addition, VF16 showed a high cytotoxicity with IC50 values of 33.52 ± 2.57, 54.63 ± 0.09, and 30.38 ± 1.37 µM against the A431, A549, and H1975 cancer cell lines, respectively. From 500-ns MD simulation, the structural stability of VF16 in complex with EGFR-TK was quite stable, suggesting that this compound could be a novel small molecule inhibitor targeting EGFR-TK.

scholarly journals Effect of Propolis Preparations on Transepithelial Electrical Potential, Resistance, and Ion Transport in In Vitro Study

2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Paulina Smyk ◽  
Iga Hołyńska-Iwan ◽  
Dorota Olszewska-Słonina
Keyword(s):  
Electrical Resistance ◽  
Ussing Chamber ◽  
Electrical Potential ◽  
In Vitro Study ◽  
Ethanol Extract ◽  
Ringer Solution ◽  
Chloride Ions ◽  
Propolis Extract ◽  
Ions Transport

Background. Propolis and its ethanol extract show positive germicidal, bacteriostatic, and anti-inflammatory antioxidants and regenerative properties after use on the surface of the skin. Propolis is in common use in production of cosmetics and in folk medicine. The influence of this resinous mixture on ion channels, channels located in skin cells membranes and skin electrical resistance, was not explained. Objective. The main aim of the study was the evaluation of electrophysiological skin parameters during mechanical and chemical-mechanical stimulation after use of ethanol extract of propolis and propolis ointment in comparison with iso-osmotic Ringer solution. Methods. Skin fragments were taken from white New Zealand rabbits and distributed into three experimental groups which were incubated in ethanol extract of propolis (EEP), propolis ointment, and Ringer solution. Then they were placed in a Ussing chamber to measure electrophysiological parameters values. Results. In this study the influence of EEP on changes in value of electrical potential during block of chloride ions transport at the same time was observed. Ethanol propolis extract dissolved in water increases the transepidermal sodium ions transport in contrast to propolis ointment. Conclusion. The way of preparation cosmetics, which contain propolis, has effects on transepidermal ions transport in the rabbit’s skin. The value of skin electrical resistance is changing with penetration depth of active propolis substances contained in cosmetics.

scholarly journals Cleavage and Inactivation of ATM during Apoptosis

1999 ◽  
Vol 19 (9) ◽  
pp. 6076-6084 ◽  
Author(s):  
Graeme C. M. Smith ◽  
Fabrizio d’adda di Fagagna ◽  
Nicholas D. Lakin ◽  
Stephen P. Jackson
Keyword(s):  
Dna Damage ◽  
Dna Binding ◽  
Caspase 3 ◽  
Cysteine Proteases ◽  
Dominant Negative ◽  
Protein Kinase Activity ◽  
Dna Damage Signalling ◽  
In Cells

ABSTRACT The activation of the cysteine proteases with aspartate specificity, termed caspases, is of fundamental importance for the execution of programmed cell death. These proteases are highly specific in their action and activate or inhibit a variety of key protein molecules in the cell. Here, we study the effect of apoptosis on the integrity of two proteins that have critical roles in DNA damage signalling, cell cycle checkpoint controls, and genome maintenance—the product of the gene defective in ataxia telangiectasia, ATM, and the related protein ATR. We find that ATM but not ATR is specifically cleaved in cells induced to undergo apoptosis by a variety of stimuli. We establish that ATM cleavage in vivo is dependent on caspases, reveal that ATM is an efficient substrate for caspase 3 but not caspase 6 in vitro, and show that the in vitro caspase 3 cleavage pattern mirrors that in cells undergoing apoptosis. Strikingly, apoptotic cleavage of ATM in vivo abrogates its protein kinase activity against p53 but has no apparent effect on the DNA binding properties of ATM. These data suggest that the cleavage of ATM during apoptosis generates a kinase-inactive protein that acts, through its DNA binding ability, in a trans-dominant-negative fashion to prevent DNA repair and DNA damage signalling.

scholarly journals THE EFFECT OF PROPOLIS EXTRACT AND PROPOLIS CANDIES ON THE GROWTH OF AGGREGATIBACTER ACTINOMYCETEMCOMITANS ATCC 43718

2017 ◽  
Vol 10 (17) ◽  
pp. 26
Author(s):  
Khodijah Khodijah ◽  
Ratna Farida ◽  
Nurtami Soedarsono
Keyword(s):  
Minimum Inhibitory Concentration ◽  
Inhibitory Concentration ◽  
Minimum Bactericidal Concentration ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Spectrophotometric Analysis ◽  
Propolis Extract ◽  
Post Exposure ◽  
Colony Forming Units

Objective: This experiment aimed to analyze the effect of propolis extract and propolis containing candies on the growth of Aggregatibacter actinomycetemcomitans using spectrophotometric analysis and colony-forming units (CFU) counts.Methods: After A. actinomycetemcomitans were exposed to propolis extract and candies, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined with spectrophotometry and post-exposure colony counting.Results: The MIC of propolis extract against A. actinomycetemcomitans was determined to be 10%, and the MBC was 20%. A decrease in the total CFU count of A. actinomycetemcomitans was observed after propolis extract and candy exposure.Conclusions: Propolis extract and propolis candies were effective in inhibiting the growth of A. actinomycetemcomitans ATCC 43718 in vitro.

scholarly journals Microfluidic deep mutational scanning of the human executioner caspases reveals differences in structure and regulation

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Hridindu Roychowdury ◽  
Philip A. Romero
Keyword(s):  
Large Scale ◽  
Cysteine Proteases ◽  
Amino Acid Substitutions ◽  
Substrate Preferences ◽  
Functional Differences ◽  
Caspase Family ◽  
Executioner Caspases ◽  
The Impact ◽  
Human Caspase

AbstractThe human caspase family comprises 12 cysteine proteases that are centrally involved in cell death and inflammation responses. The members of this family have conserved sequences and structures, highly similar enzymatic activities and substrate preferences, and overlapping physiological roles. In this paper, we present a deep mutational scan of the executioner caspases CASP3 and CASP7 to dissect differences in their structure, function, and regulation. Our approach leverages high-throughput microfluidic screening to analyze hundreds of thousands of caspase variants in tightly controlled in vitro reactions. The resulting data provides a large-scale and unbiased view of the impact of amino acid substitutions on the proteolytic activity of CASP3 and CASP7. We use this data to pinpoint key functional differences between CASP3 and CASP7, including a secondary internal cleavage site, CASP7 Q196 that is not present in CASP3. Our results will open avenues for inquiry in caspase function and regulation that could potentially inform the development of future caspase-specific therapeutics.

Sign in / Sign up

Export Citation Format

Share Document