scholarly journals Fluorescence-based determination of enzyme activity of recombinant CYP51b1 (sterol 14α-demethylase) with coumarin derivatives

2010 ◽  
Vol 56 (1) ◽  
pp. 132-137 ◽  
Author(s):  
N.A. Petushkova ◽  
A.V. Lisitsa ◽  
V.F. Pozdnev ◽  
I.I. Karuzina
Keyword(s):  
Enzyme Activity ◽  
Acetic Acid ◽  
Electron Donor ◽  
Model System ◽  
Coumarin Derivatives ◽  
Current Investigation ◽  
Soluble Model ◽  
Highly Sensitive

The current investigation was undertaken with the aim to carry out an in vitro evaluation of the ability of coumarin derivatives as probe substrates to predict the activity of CYP51b1. The results obtained indicate that 7-aminocoumarin-4-acetic acid (ACAC) can be used to determine the recombinant CYP51b1 activity. Determination of CYP51b1 activity with ACAC is based on the direct registration of fluorescence increasing at 30°C. It was found also that BMR in a simple soluble model system can be used as an electron donor for CYP51B1. Fluorescence-based assay is highly sensitive and can be used for the screening of sterol 14alpha-demethylase inhibitors.

scholarly journals Influence of Indole acetic acid and Tryptophan on production of Vinblastine and Vincristine of Catharanthus roseous callus cells in the accumulation media of In Vitro tissue culture

2010 ◽  
Vol 7 (3) ◽  
pp. 1113-1119
Author(s):  
Baghdad Science Journal
Keyword(s):  
Tissue Culture ◽  
High Performance Liquid Chromatography ◽  
Acetic Acid ◽  
Liquid Chromatography ◽  
In Vitro Culture ◽  
High Performance ◽  
Indole Acetic Acid ◽  
Culture Technique

This study on the plant of Ain –AL Bason Catharanthus roseous showed the ability of callus cells that is produced by In Vitro culture technique and transformed to the accumulated media (MS 40gm/L sucrose ,2gm/L IAA Indole acetic acid , 0.5gm/L Tryptophan) to produce Vinblastine and Vincristine compounds. Extraction, purification and quantitive determination of Vinblastine and Vincristine compounds using High performance liquid chromatography technique (HPLC)were carried out. The results showed that the highest concentration of Vinblastine and Vincristine compounds were ( 4.653,12.5 (ppm /0.5 dry Wight respectively from transformed callus cells from MS 40 gm /L sucrose , 2 gm / L NAA Naphthaline acetic acid .

scholarly journals Comparative Analysis of Therapeutically Important Indole Compounds in in vitro Cultures of Hypericum perforatum Cultivars by HPLC and TLC Analysis Coupled with Densitometric Detection

2014 ◽  
Vol 9 (10) ◽  
pp. 1934578X1400901 ◽  
Author(s):  
Bożena Muszyńska ◽  
Halina Ekiert ◽  
Inga Kwiecień ◽  
Anna Maślanka ◽  
Rawad Zodi ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Hypericum Perforatum ◽  
Dry Mass ◽  
In Vitro Cultures ◽  
Indole Compounds ◽  
Densitometric Detection ◽  
Tlc Analysis

Five indole compounds (5-hydroxy-L-tryptophan, L-tryptophan, indole-3-acetic acid, melatonin, serotonin) and hypericin were identified and quantified in methanolic extracts of shoot cultures of three Hypericum perforatum cultivars (Helos, Elixir, Topas) growing on two variants of Murashige -Skoog medium differing in concentrations of growth regulators (naphthalene-1-acetic acid and 6-benzylaminopurine). Extracts of the aboveground parts of field-grown plants ( Hyperici herba) were also analyzed by HPLC and TLC analysis coupled with densitometric detection. Determination of four compounds was based on our assay described earlier. The methods of determination of 5-hydroxy-L-tryptophan and hypericin were developed and validated in this study. The composition and contents of the metabolites under study differed between the cultivars cultured in vitro and between medium variants containing diverse contents of growth regulators. The contents of individual indole compounds in the biomass from in vitro cultures ranged from 39.6 to 343.2 mg/100 g dry mass. 5-Hydroxy-L-tryptophan was the dominating metabolite (from 78.2 to 343.2 mg/100 g dry mass). Extracts from shoots of the cultivar Helos also contained high contents of serotonin (319.9 and 197.4 mg/100 g dry mass). The contents of indole compounds in Hyperici herba were also diverse (from 7.1 to 55.3 mg/100 g dry mass). 5-Hydroxy-L-tryptophan was the dominating metabolite as well. Hypericin content of Hyperici herba, equaling 12.2 mg/100 g dry mass was from 3.3 to 10 times higher than in extracts from shoots cultured in vitro. The present report is the first analysis of endogenous accumulation of indole compounds in Hyperici herba which involves, apart from melatonin, four other compounds.

scholarly journals Utilization of a new gold/Schiff-base iron(iii) complex composite as a highly sensitive voltammetric sensor for determination of epinephrine in the presence of ascorbic acid

RSC Advances ◽  
2016 ◽  
Vol 6 (104) ◽  
pp. 101888-101899 ◽  
Author(s):  
Adam Gorczyński ◽  
Maciej Kubicki ◽  
Klaudia Szymkowiak ◽  
Teresa Łuczak ◽  
Violetta Patroniak
Keyword(s):  
Ascorbic Acid ◽  
Schiff Base ◽  
Schiff Base Complex ◽  
Voltammetric Sensor ◽  
Base Complex ◽  
Highly Sensitive

A new voltammetric sensor based on an iron(iii) Schiff-base complex/Au composite is synthesized and applied for the in vitro detection of epinephrine.

scholarly journals Highly Sensitive Micellar Enhanced Spectrofluorimetric Method for Determination of Mirtazapine in Tablets and Human Urine: Application to In Vitro Drug Release and Content Uniformity Test

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Hany W. Darwish ◽  
Ahmed H. Bakheit ◽  
Raed M. Alharbi
Keyword(s):  
Drug Release ◽  
Human Urine ◽  
Dosage Form ◽  
Content Uniformity ◽  
In Vitro Drug Release ◽  
Spectrofluorimetric Method ◽  
Highly Sensitive ◽  
Spiked Human Urine

A highly sensitive and simple micelle enhanced spectrofluorimetric method was developed for assaying mirtazapine (MRZ) in REMERON® tablets and spiked human urine directly without the need of derivatizing agent. The basis of the current procedure is the examination of the relative fluorescence intensity (RFI) of MRZ in sodium lauryl sulphate (SLS) micellar medium. The RFI of MRZ in water was enhanced markedly on addition of SLS. The RFI was measured at 403 nm after excitation at 320 nm. The fluorescence-concentration relationship was linear over the range 1–500 ng/mL, with lower detection limit of 0.399 ng/mL. The proposed method was successfully applied to the determination of MRZ in dosage form and spiked human urine. Recovery percentages of MRZ utilizing the current method were99.05±1.83,98.37±1.96, and100.41±2.61% for pure powder, pharmaceutical dosage form, and spiked human urine, respectively. The application of the proposed method was extended to test content uniformity and the in vitro drug release of REMERON tablets, according to USP guidelines.

Ein chromatographisches Verfahren zur Bestimmung typenspezifischer Poliovirus-Antikörper mit 32P-markiertem Polioviru

1963 ◽  
Vol 18 (10) ◽  
pp. 798-808 ◽  
Author(s):  
Reiner Thomssen
Keyword(s):  
Aluminium Hydroxide ◽  
Virus Antibody ◽  
Poliovirus Type ◽  
Specific Antibodies ◽  
In Vitro Technique ◽  
Step Procedure ◽  
Highly Sensitive

A highly sensitive in-vitro technique for determination of homologous type-specific antibodies against polioviruses is described. 32P-labelled poliovirus type 1. strain Mahoney, purified by a three-step-procedure (15 000 rpm, aluminiumhydroxide. Ecteolacellulose) is mixed with homologous type-specific antiserum. After incubation the mixtures are adsorbed to aluminium hydroxide. Virus-antibody-com-plexes are more firmly bound to this adsorbent than virus without antibody: the distinguishing parameter is the concentration of divalent phosphate in the adsorption or the elution fluid. The firmness of binding of virus-antibody-complexes to aluminium hydroxide is further a function of the dilution of antiserum. This leads to a suitable end-point for reading antiserum titers. There is a good correlation between titers determined by this method and titers, determined by common neutralization tests. The method may be used to judge the virus-specifity of radioactivity after purification of 32P-labelled poliovirus.

scholarly journals The kinetic characteristics of human and trypanosomatid phosphofructokinases for the reverse reaction

2019 ◽  
Vol 476 (2) ◽  
pp. 179-191 ◽  
Author(s):  
Peter M. Fernandes ◽  
James Kinkead ◽  
Iain W. McNae ◽  
Frédéric Bringaud ◽  
Paul A.M. Michels ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Reverse Reaction ◽  
Kinetic Properties ◽  
Kinetic Characteristics ◽  
Inorganic Pyrophosphate ◽  
Physiological Relevance ◽  
Fructose 1,6 Bisphosphate

Abstract Eukaryotic ATP-dependent phosphofructokinases (PFKs) are often considered unidirectional enzymes catalysing the transfer of a phospho moiety from ATP to fructose 6-phosphate to produce ADP and fructose 1,6-bisphosphate. The reverse reaction is not generally considered to occur under normal conditions and has never been demonstrated for any eukaryotic ATP-dependent PFKs, though it does occur in inorganic pyrophosphate-dependent PFKs and has been experimentally shown for bacterial ATP-dependent PFKs. The evidence is provided via two orthogonal assays that all three human PFK isoforms can catalyse the reverse reaction in vitro, allowing determination of kinetic properties. Additionally, the reverse reaction was shown possible for PFKs from three clinically important trypanosomatids; these enzymes are contained within glycosomes in vivo. This compartmentalisation may facilitate reversal, given the potential for trypanosomatids to have an altered ATP/ADP ratio in glycosomes compared with the cytosol. The kinetic properties of each trypanosomatid PFK were determined, including the response to natural and artificial modulators of enzyme activity. The possible physiological relevance of the reverse reaction in trypanosomatid and human PFKs is discussed.

scholarly journals 2-{[(4-Hydroxy-3,5-dimethoxyphenyl)methylidene]hydrazinylidene}-4-oxo-1,3-thiazolidin-5-yl Acetic Acid

Molbank ◽  
10.3390/m1009 ◽  
2018 ◽  
Vol 2018 (3) ◽  
pp. M1009 ◽  
Author(s):  
Sangeetha Karanth ◽  
Badiadka Narayana ◽  
Sharath Kodandoor ◽  
Balladka Sarojini
Keyword(s):  
Acetic Acid ◽  
Title Compound ◽  
Radical Scavenging ◽  
Antioxidant Property ◽  
Mass Spectral ◽  
Bacterial Enzyme ◽  
Ft Ir ◽  
Antibacterial And Antifungal

Thia-Michael addition of 2-[(4-hydroxy-3,5-dimethoxyphenyl)methylidene]hydrazine-1-carbothioamide (1) with maleic anhydride results in the formation of the title compound 2-{[(4-hydroxy-3,5-dimethoxyphenyl)methylidene]hydrazinylidene}-4-oxo-1,3-thiazolidin-5-yl acetic acid 2. The precursor 1 is synthesized by the reaction of 4-hydroxy-3,5-dimethoxybenzaldehyde and thiosemicarbazide in the presence of glacial acetic acid as the catalyst. The structure of the title compound is determined by elemental analysis, FT-IR, 1H-NMR, 13C-NMR and mass spectral data. In order to determine the molecular interactions with the bacterial enzyme, the title compound is further docked into the active site of the MurB protein of Staphylococcus aureus (PDB ID: 1HSK). The in vitro antibacterial and antifungal activity of the title compound is carried out in order to appraise its antimicrobial efficacy by determination of zone of inhibition and minimal inhibitory concentration. The compound is also evaluated for its antioxidant property by 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging assay.

scholarly journals Simultaneous Spectrophotometric Determination of Chromium(VI) and Vanadium(V) by using 3,4-Dihydroxybenzaldehyde isonicotinoyl hydrazone (3,4-DHBINH)

2009 ◽  
Vol 6 (s1) ◽  
pp. S459-S465 ◽  
Author(s):  
Lakshmi Narayana ◽  
Suvarapu ◽  
Adinarayana Reddy Somala ◽  
Prathima Bobbala ◽  
Hwang Inseong ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Metal Ions ◽  
Sodium Acetate ◽  
Atomic Absorption Spectrophotometry ◽  
Photometric Method ◽  
Sodium Acetate Buffer ◽  
Absorption Spectrophotometry ◽  
Chromium Vi ◽  
Highly Sensitive

A simple, rapid, highly sensitive and new simultaneous spectro- photometric method is proposed for the analysis of chromium(VI) and vanadium(V) without separation by using 3,4-dihydroxybeznaldehyde isonicotinoyl hydrazone (3,4-DHBINH). The reagent reacts with the two metal ions in acetic acid-sodium acetate buffer of pH 5.5 and gives maximum absorbances at 400 nm and 360 nm for chromium(VI) and vanadium(V), respectively. Both the metal ions gives 1: 1 (M:L) complexes with the reagent. Effect of various diverse ions also studied. The instability constants for the two complexes were also evaluated. This method was successfully applied for the determination of chromium(VI) and vanadium(V) in various spiked samples. The validity of the method was checked by comparing with the results obtained by atomic absorption spectrophotometry.

scholarly journals In vitro Anti-Arthritic activity of Pseudarthria viscida against Protein Denaturation and Proteinase enzyme

2020 ◽  
Vol 11 (2) ◽  
pp. 284-288
Author(s):  
Pandian P
Keyword(s):  
Enzyme Activity ◽  
Aqueous Extract ◽  
Protein Denaturation ◽  
Bone Erosion ◽  
Diclofenac Sodium ◽  
Ethanol Extract ◽  
Maximum Inhibition

Arthritis is an autoimmune with chronic inflammatory, the patient has very painful due deformities and bone erosion which is caused by damage of the joints. The plant Pseudarthria viscida was collected from the Thirunelveli district and extracted with aqueous and ethanol solvent. The two method was used for determination of invitro anti-arthritic activity. The Inhibition of Protein Denaturation Method shows the anti-arthritic activity with the value from 40.46±0.72 to 78.36±0.64 for aqueous extract and 48.62±0.86 to 84.42±0.86 for ethanol extract and Inhibition of Proteinase Enzyme Activity shows 38.62±0.32 to 72.58±0.58 in aqueous extract and 46.28±0.58 to 80.52±0.56 in ethanol extract. Diclofenac sodium were used as standard, the concentration is 100, 200, 300, 400, and 500. In both the method the concentration of 500Microgram per milliliters shows maximum inhibition and compare to both extract the ethanol shows better activity than aqueous extract.

Validation of a Real-Time ISE Methodology to Quantify the Influence of Inhibitors of Demineralization Kinetics in vitro Using a Hydroxyapatite Model System

2018 ◽  
Vol 52 (6) ◽  
pp. 598-603
Author(s):  
Wei-Te Huang ◽  
Saroash Shahid ◽  
Paul Anderson
Keyword(s):  
Acetic Acid ◽  
Real Time ◽  
Model System ◽  
Ion Selective Electrodes ◽  
Percentage Reduction ◽  
Inhibitor Concentration ◽  
Rate Of Increase ◽  
Log Linear ◽  
Calcium Loss

The aim was to validate a novel protocol to measure the cariostatic efficacies of demineralization inhibitors by repeating previous SMR (scanning microradiography) studies investigating the dose response of Zn2+ and F– on demineralization kinetics in vitro using real-time Ca2+ ion selective electrodes (ISEs). In this study, Ca2+ release was used as a proxy for the extent of demineralization. Forty-eight hydroxyapatite (HAP) discs were allocated into 16 groups (n = 3) and adding either increasing [Zn2+], or [F–], similar to those used in the previous SMR studies. Each HAP disc was immersed in 50 mL, pH 4.0, buffered acetic acid for 1 h, and real-time ISE methodology was used to monitor the rate of increase in [Ca2+] in the demineralization solution. Next, either zinc acetate or sodium fluoride was added into each demineralization solution accordingly. Then after each [Zn2+] or [F–] addition, the HAP disc was further demineralized for 1 h, and ISE measurements were continued. The percentage reduction in the rate of calcium loss from hydroxyapatite (PRCLHAP) at each [Zn2+] or [F–] was calculated from the decrease in Ca2+ release rate, similar to that used in the previous SMR studies. A log-linear relationship between mean PRCLHAP and inhibitor concentration was found for both Zn2+ and F–, similar to that reported for each ion in the previous SMR studies. In conclusion, real-time Ca2+ ISE systems can be used to measure the cariostatic efficacies of demineralization inhibitors.

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