scholarly journals Assimilation of Cholesterol by Monascus purpureus

2020 ◽  
Vol 6 (4) ◽  
pp. 352
Author(s):  
Theresa P. T. Nguyen ◽  
Margaret A. Garrahan ◽  
Sabrina A. Nance ◽  
Catherine E. Seeger ◽  
Christian Wong
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Cholesterol Biosynthesis ◽  
Cholesterol Content ◽  
Monascus Purpureus ◽  
Growth Media ◽  
Red Yeast Rice ◽  
Cholesterol Reduction ◽  
Statin Drugs

Monascus purpureus, a filamentous fungus known for its fermentation of red yeast rice, produces the metabolite monacolin K used in statin drugs to inhibit cholesterol biosynthesis. In this study, we show that active cultures of M. purpureus CBS 109.07, independent of secondary metabolites, use the mechanism of cholesterol assimilation to lower cholesterol in vitro. We describe collection, extraction, and gas chromatography-flame ionized detection (GC-FID) methods to quantify the levels of cholesterol remaining after incubation of M. purpureus CBS 109.07 with exogenous cholesterol. Our findings demonstrate that active growing M. purpureus CBS 109.07 can assimilate cholesterol, removing 36.38% of cholesterol after 48 h of incubation at 37 °C. The removal of cholesterol by resting or dead M. purpureus CBS 109.07 was not significant, with cholesterol reduction ranging from 2.75–9.27% throughout a 72 h incubation. Cholesterol was also not shown to be catabolized as a carbon source. Resting cultures transferred from buffer to growth media were able to reactivate, and increases in cholesterol assimilation and growth were observed. In growing and resting phases at 24 and 72 h, the production of the mycotoxin citrinin was quantified via high-performance liquid chromatography-ultraviolet (HPLC-UV) and found to be below the limit of detection. The results indicate that M. purpureus CBS 109.07 can reduce cholesterol content in vitro and may have a potential application in probiotics.

scholarly journals Determination of Tedizolid in Bacterial Growth Medium Mueller-Hinton Broth by High-Performance Liquid Chromatography and Its Application to an In Vitro Study in the Hollow-Fiber Infection Model

Separations ◽  
2021 ◽  
Vol 8 (9) ◽  
pp. 141
Author(s):  
Khalid Iqbal ◽  
Aliki Milioudi ◽  
Elena Haro Martínez ◽  
Sebastian Georg Wicha
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Growth Medium ◽  
Bacterial Growth ◽  
High Performance ◽  
Infection Model ◽  
Growth Media ◽  
Mueller Hinton ◽  
Mueller Hinton Broth

Pharmacokinetic/pharmacodynamic (PKPD) studies of anti-infectives are frequently performed in in vitro infection models where accurate quantification of antibiotic concentrations in bacterial growth media is crucial to establish PK/PD relationships. Here, a sensitive and rapid high-performance liquid chromatography (HPLC) method was developed to quantify tedizolid (TDZ) in the bacterial growth medium Mueller-Hinton broth (MHB). Matrix components were separated by direct protein precipitation with methanol (1:1). The chromatographic separation was carried out in a Dionex Ultimate 3000 HPLC system using an Accucore® C-18 RPMS HPLC column (2.6 µm, 100 × 2.1 mm) using isocratic elution with 25% acetonitrile and 75% of 0.1% formic acid. The lower limit of quantification was 0.03 mg/L when measured at 300 nm. Following relevant European Medicine Agency guidelines, the method was successfully validated for linearity, selectivity, recovery, inter- and intra-day precision, and accuracy and stability. When applied to in vitro PKPD studies, the method successfully quantified a range of TDZ concentration (Cmin, 0.09-Cmax, 0.65 mg/L) in MHB. The analyzed concentrations were in line with the planned PK profiles. The application of the developed method to quantify TDZ in MHB in in vitro PKPD studies is warranted.

scholarly journals PHYTOCHEMICAL ANALYSIS OF CALLUS TWO VARIETIES ORTHOSIPHON ARISTATUS (BLUME) MIQ ON MURASHIGE AND SKOOG MEDIA: A STRATEGIC STEP OF SECONDARY METABOLITE PRODUCTION

2021 ◽  
pp. 71-77
Author(s):  
FAHRAUK FARAMAYUDA ◽  
TOTIK SRI MARIANI ◽  
ELFAHMI ◽  
SUKRASNO
Keyword(s):  
Secondary Metabolites ◽  
Rosmarinic Acid ◽  
High Performance ◽  
Leaf Explants ◽  
Gradient Elution ◽  
Growth Media ◽  
Phytochemical Analysis ◽  
New Information ◽  
Sterile Leaf

Objective: The research aimed to provide new information regarding the secondary metabolites content of purple and white-purple Orthosiphon aristatus (Blume) Miq. callus, which can then be used as a basis for developing towards cell suspension and ultimately producing secondary metabolites using bioreactors. Methods: Callus induction of two varieties of O. aristatus were performed by inoculating sterile leaf explants grown on Murashige and Skoog basal media supplemented with 2,4-dichlorophenoxyacetis acid 0.4 ppm. The secondary metabolites were analysed and quantified using high-performance liquid chromatography with gradient elution. Results: The results showed the growth of callus two varieties of O. aristatus in growth media MS with 2,4-D 0.4 ppm. Rosmarinic acid content in the acetone extract of the purple variety callus was 1.28% w/w, and the white-purple variety was 2.22% w/w. Conclusion: This study could form the basis for the development of rosmarinic acid production by In vitro culture modification.

Identification of a non-covalent oxytocin/neurophysin-I complex in the bovine ovary

1991 ◽  
Vol 128 (2) ◽  
pp. 305-314 ◽  
Author(s):  
L. Shukovski ◽  
J. K. Findlay ◽  
A. I. Smith
Keyword(s):  
Granulosa Cells ◽  
High Performance ◽  
Limit Of Detection ◽  
Secretory Granules ◽  
Active Form ◽  
Biologically Active ◽  
Corpora Lutea ◽  
Free Peptide ◽  
Second Peak

ABSTRACT Acid extracts of bovine preovulatory granulosa cells and corpora lutea (CL) were subjected to high-performance liquid chromatography (HPLC) and found to contain two peaks of immunoreactive (ir) oxytocin (OT), one corresponding to authentic OT and the second eluting 8 min later. The second peak was more abundant than authentic irOT in preovulatory follicles and in the early CL, but became less abundant as the CL matured (mid luteal) and was close to the limit of detection in the late CL. This peak could be detected only by an OT antiserum which recognized both the biologically active form of OT, as well as the post-translational processing intermediate Gly10-extended oxytocin. A second more specific OT antiserum (OT-933) did not recognize the second peak as strongly. Further analysis of the second peak revealed a complex of OT bound to its neurophysin (NP-I) which could be dissociated under denaturing conditions. Furthermore, we were able to create this complex in vitro by combining the two materials together under acid conditions, similar to the pH predicted in secretory granules, but not under neutral conditions. Measuring irNP-I by radioimmunoassay showed a single peak with a similar retention time to the OT/NP-I complex, confirming the identity of the unknown peak. Incubation of CL slices in culture showed a time-related release of both OT and NP-I, with OT having a greater rate of release in the mid luteal CL. These data suggest the presence of an OT/NP-I complex in the bovine preovulatory granulosa cells and CL, as well as the unbound peptide presumably within the secretory granules. The ratio of OT/NP-I complex and free peptide changes with ageing of the the CL, perhaps indicating regulated differences in the post-translational processing of the prohormone. Journal of Endocrinology (1991) 128, 305–314

scholarly journals PROTEIN BINDING STUDY OF FELODIPINE USING VALIDATED LIQUID CHROMATOGRAPHIC METHOD

2017 ◽  
pp. 134-139
Author(s):  
Swarna Vijitha ◽  
P. Rajavel ◽  
K. Udayasree
Keyword(s):  
Protein Binding ◽  
High Performance ◽  
Limit Of Detection ◽  
Binding Studies ◽  
Limit Of Quantification ◽  
Permeable Membrane ◽  
Percentage Recovery ◽  
Ich Guidelines ◽  
Liquid Chromatographic

Objective: The aim of present study was to develop and validate a new simple, easy, selective, precise, accurate reverse phase high-performance liquid chromatography for the estimation of felodipine in bulk and pharmaceutical dosage form.Methods: The separation was carried on HPLC system consisting C18 column (150 mm ×4.6 nm, 5 µm) at room temperature coupled with a phenomenixcolumn silica with flow rate 1 ml/min. The mobile phase used was methanol: acetonitrile in the ratio of 50: 50. The drug was detected using UV-visible detector at the wavelength of 230 nm and run time was 10 min.Results: The retention time was 3.138 min. Linearity was observed in the concentration range of 5-25μg/ml. The accuracy of the method was assessed by percentage recovery studies at three different levels at 80%, 100% and 120% of its working concentration. The percentage recovery of felodipine in the developed method was found to be in the ranges of from 99.81-100.00% that indicates the good accuracy of the method. The percentage % RSD of precision was found to be less than 2%. The method was validated as per ICH guidelines. The developed method was employed in in vitro protein binding studies using semi permeable membrane and performed by plotting calibration curve (peak area vs concentration) the % drug release of felodipine was calculated.Conclusion: The proposed method was found to be simple, precise, accurate and consistent. The validated parameters are statistically validated for linearity, precision and limit of detection, limit of quantification, robustness, ruggedness were concluded. 

scholarly journals In Vitro and In Vivo Correlation of Colon-Targeted Compression-Coated Tablets

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Siddhartha Maity ◽  
Amit Kundu ◽  
Sanmoy Karmakar ◽  
Biswanath Sa
Keyword(s):  
Drug Release ◽  
Drug Absorption ◽  
High Performance ◽  
Limit Of Detection ◽  
High Performance Liquid Chromatographic ◽  
Absorption Study ◽  
Limit Of Quantification ◽  
In Vitro Drug Release

This study was performed to assess and correlate in vitro drug release with in vivo absorption of prednisolone (PDL) from a colon-targeted tablet prepared by compression coating of core tablet. In vivo drug absorption study was conducted using a high performance liquid chromatographic (HPLC) method, which was developed and validated for the estimation of PDL in rabbit plasma. The calibration curve showed linearity in the concentration range of 0.05 to 50 μg/mL with the correlation coefficient (r) of 0.999. The method was specific and sensitive with the limit of detection (LOD) and lower limit of quantification (LLOQ) of 31.89±1.10 ng/mL and 96.63±3.32 ng/mL, respectively. The extraction recovery (ER) of PDL from three different levels of quality control (QC) samples ranged from 98.18% to 103.54%. In vitro drug release study revealed that less than 10% drug was released in 6.34 h and almost complete (98.64%) drug release was achieved in the following 6 h. In vivo drug absorption study demonstrated lower values of Cmax, AUCtotal, and protracted Tmax from compression-coated tablet. The results confirmed the maximum release of drug in the colon while minimizing release in the upper gastrointestinal tract (GIT). An excellent in vitro and in vivo correlation (IVIVC) was also achieved after considering the lag time.

scholarly journals Development and Validation of RP-HPLC-PDA Method for the Analysis of Diclofenac Sodium in the In Vitro Transdermal Permeation Samples

2019 ◽  
Vol 9 (2) ◽  
pp. 212-216 ◽  
Author(s):  
Saisrianusha Valluru ◽  
Buchi N Nalluri
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Skin Permeation ◽  
Diclofenac Sodium ◽  
Hplc Method ◽  
Limit Of Quantification ◽  
Validation Parameters ◽  
In Vitro Skin Permeation ◽  
Development And Validation

A new analytical method using high-performance liquid chromatography coupled with photo diode array detection was developed and validated for the quantification of Diclofenac (DIC) from in vitro skin permeation samples. Analysis was performed using a Phenomenex C18 column (150 x 4.6mm, 5µm) with 10mM ammonium acetate: Acetonitrile (62:38% v/v) as the mobile phase in isocratic mode and eluents were monitored at 276nm. DIC was eluted at 3.1min and showed a good linearity in the concentration range of 0.2-3µg/mL with a correlation coefficient >0.999. The validation parameters, such as specificity, linearity, accuracy and limit of detection, limit of quantification, precision, robustness fulfilled the regulatory requirements. The developed HPLC method was successfully used for the analysis of DIC in samples obtained from transdermal diffusate samples.

scholarly journals Oncogenic role of SOX9-DHCR24-cholesterol biosynthesis axis in IGH-BCL2 positive diffuse large B-cell lymphomas

Blood ◽  
2021 ◽  
Author(s):  
Yajie Shen ◽  
Jingqi Zhou ◽  
Kui Nie ◽  
Shuhua Cheng ◽  
Zhengming Chen ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Line ◽  
Cholesterol Synthesis ◽  
Cholesterol Biosynthesis ◽  
Cholesterol Content ◽  
B Cell Lymphomas ◽  
Tumor Load ◽  
Large B Cell

Although oncogenicity of the stem cell regulator SOX9 has been implicated in many solid tumors, its role in lymphomagenesis remains largely unknown. In this study, we showed that SOX9 is overexpressed preferentially in a subset of diffuse large B-cell lymphomas (DLBCL) harboring IGH-BCL2 translocations. SOX9 positivity in DLBCL correlates with advanced stage of disease. Silencing of SOX9 decreased cell proliferation, induced G1/S arrest and increased apoptosis of DLBCL cells, both in vitro and in vivo. Whole transcriptome analysis and CHIP-seq assays identified DHCR24, a terminal enzyme in cholesterol biosynthesis, as a direct target of SOX9, which promotes cholesterol synthesis by increasing DHCR24 expression. Enforced expression of DHCR24 was capable of rescuing the phenotypes associated with SOX9 knockdown in DLBCL cells. In DLBCL cell line xenograft models, SOX9 knockdown resulted in lower DHCR24 level, reduced cholesterol content and decreased tumor load. Pharmacological inhibition of cholesterol synthesis also inhibited DLBCL xenograft tumorigenesis, the reduction of which is more pronounced in DLBCL cell line with higher SOX9 expression, suggesting that it may be addicted to cholesterol. In summary, our study demonstrates that SOX9 can drive lymphomagenesis through DHCR24 and the cholesterol biosynthesis pathway. This SOX9-DHCR24-cholesterol biosynthesis axis may serve as a novel treatment target for DLBCL.

scholarly journals High-Performance Liquid Chromatographic Ultraviolet Determination of Memantine Hydrochloride afterIn VitroTransdermal Diffusion Studies

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Sergio del Rio-Sancho ◽  
César E. Serna-Jiménez ◽  
M. Aracely Calatayud-Pascual ◽  
Cristina Balaguer-Fernández ◽  
Andrés Femenía-Font ◽  
...  
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Limit Of Quantification ◽  
Detection Response ◽  
Uv Visible ◽  
Liquid Chromatographic

The purpose of the present work was to validate an accurate and precise high-performance liquid chromatography (HPLC) method involving ultraviolet detection for the quantitative analysis of memantine hydrochloride. In order to analyze a molecule with no chromophoric groups that could be detected by a UV/visible detector, it was necessary to extract the drug and to perform a dansylation reaction that enabled the UV/visible detection of the derivatized molecule. Separation was carried out with a 150 mm Kromasil C18 column at room temperature. The detection response, at 218 nm, was found to be linear in the concentration range from 0.5 to 50 μg/mL. The method was validated for specificity, linearity, precision, accuracy, limit of detection, limit of quantification, and robustness. The limit of detection (LOD) was 0.144 μg/mL, and the limit of quantification (LOQ) was 0.437 μg/mL. The dansylated memantine complex was stable for at least five days in all the conditions evaluated. The potential use of this method has been demonstrated by the quantification of memantine hydrochloride contained in samples from the study of itsin vitrotransdermal permeation.

scholarly journals Selenium and Zinc Biofortification of Pleurotus eryngii Mycelium and Fruiting Bodies as a Tool for Controlling Their Biological Activity

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 889 ◽  
Author(s):  
Piotr Zięba ◽  
Katarzyna Kała ◽  
Anna Włodarczyk ◽  
Agnieszka Szewczyk ◽  
Edward Kunicki ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Hplc Method ◽  
Pleurotus Eryngii ◽  
Array Detector ◽  
Growth Media ◽  
Fruiting Bodies ◽  
Cultivated Mushroom

Pleurotus eryngii (DC:Fr.) Quel. is a cultivated mushroom of high culinary value and medicinal properties. Mycelium of P. eryngii is characterized by the ability of effective bio-elements absorption from growth media so it could be biofortified with trace elements with a functional activity in the human body. In this study, the ability of P. eryngii mycelia from in vitro cultures as well as fruiting bodies were investigated in terms of their effectiveness in zinc and selenium accumulation. The effect of Se and Zn biofortification on productivity, chemical compounds, and bio-elements content of P. eryngii was determined as well. To enhance Se and Zn content in P. eryngii fruiting bodies and mycelia, substrates were supplemented with sodium selenite, at a concentration of 50 mg L−1, zinc sulfate, and zinc hydro-aspartate at a concentration of 87.2 and 100.0 mg L−1, respectively. Mentioned Zn concentrations contained the same amount of zinc(II) ions, namely 20 mg L−1. The content of organic compounds include phenolic compounds and lovastatin, which were determined by a high-performance liquid chromatography with diode-array detector (HPLC-DAD) and reverse phase high-performance liquid chromatography (RP-HPLC) method with UV detection. The ability of P. eryngii to accumulate zinc and selenium from the culture medium was demonstrated. The degree of accumulation of zinc turned out to be different depending on the type of salt used. The present study also showed that conducting mycelium of P. eryngii in in vitro culture, with a higher content of zinc ions, can result in obtaining the materials with better antioxidant ability. The results of this study can be used to develop the composition of growing media, which ensures the production of biomass with the desired composition of elements.

The Effect of Oxidized Cholesterol on the Aorta of Rabbits- Scanning Electron Microscopic Observations

1982 ◽  
Vol 40 ◽  
pp. 340-341
Author(s):  
S. K. Pena ◽  
C. B. Taylor ◽  
J. Hill ◽  
J. Safarik
Keyword(s):  
Cholesterol Biosynthesis ◽  
Cholesterol Content ◽  
Arterial Injury ◽  
Electron Microscopic ◽  
Aortic Smooth Muscle ◽  
Oxidized Cholesterol ◽  
Initial Event ◽  
Membrane Structure And Function ◽  
Scanning Electron Microscopic ◽  
And Function

Introduction: Oxidized cholesterol derivatives have been demonstrated in various cell cultures to be very potent inhibitors of 3-hvdroxy-3- methylglutaryl Coenzyme A reductase which is a principle regulator of cholesterol biosynthesis in the cell. The cholesterol content in the cells exposed to oxidized cholesterol was found to be markedly decreased. In aortic smooth muscle cells, the potency of this effect was closely related to the cytotoxicity of each derivative. Furthermore, due to the similarity of their molecular structure to that of cholesterol, these oxidized cholesterol derivatives might insert themselves into the cell membrane, alter membrane structure and function and eventually cause cell death. Arterial injury has been shown to be the initial event of atherosclerosis.

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