Coronatine production in vitro and in vivo and its relation to symptom development in bacterial blight of soybean

1982 ◽  
Vol 60 (5) ◽  
pp. 645-650 ◽  
Author(s):  
S. S. Gnanamanickam ◽  
A. N. Starratt ◽  
E. W. B. Ward
Keyword(s):  
Bacterial Blight ◽  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Culture Filtrates ◽  
Soybean Plants ◽  
Systemic Symptoms ◽  
Soybean Leaves

Coronatine was detected in culture filtrates of 12 out of 19 pathogenic strains of Pseudomonas glycinea examined, but not in culture filtrates of strains of P. phaseolicola, P. syringae, or P. tabaci. A reversed-phase high-performance liquid chromatographic procedure was developed for coronatine quantitation. Generally, coronatine production by strains in culture correlated with their ability to induce systemic symptoms (chlorosis and stunting) in inoculated soybean plants. Application of purified preparations of coronatine to unifoliate leaves of soybean plants resulted in localized chlorosis, development of chlorosis in subsequently developing trifoliate leaves, and stunting of plant growth, similar to symptoms induced by infection. Coronatine was demonstrated in soybean leaves infected with P. glycinea but was not detected in healthy leaves. The results indicate that coronatine can play an important role in the development of symptoms of bacterial blight of soybean, but the demonstration that some pathogenic strains do not produce coronatine indicates that it may not be essential for pathogenicity.

The effects of BR-DIM (BioResponse 3, 3’-Diindolylmethane) administered pre-prostatectomy on the androgen receptor (AR).

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 1560-1560
Author(s):  
Elisabeth I. Heath ◽  
Clara Hwang ◽  
Michael L. Cher ◽  
Lance K. Heilbrun ◽  
Isaac Powell ◽  
...  
Keyword(s):  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Primary Objective ◽  
Prostate Tissue ◽  
Cruciferous Vegetables ◽  
Nuclear Exclusion ◽  
Tandem Mass Spectrometric ◽  
Tissue Specimens

1560 Background: Consumption of cruciferous vegetables is associated with decreased risk of prostate cancer (PCa). 3,3’-diindolylmethane (DIM), an in vivo active compound formed after consumption of cruciferous vegetables, down-regulates the AR, and causes its nuclear exclusion which results in in vitro growth arrest and apoptosis of PCa cells. We conducted a biomarker trial evaluating BR-DIM in pre-prostatectomy patients with the primary objective of measuring DIM levels in prostate tissue and in plasma. Methods: Patients with organ-confined PCa who were candidates for surgery were treated with BR-DIM at a dose of 225 mg orally twice daily for a minimum of 14 days. Patients did not receive any androgen deprivation therapy. Patients received their last dose of BR-DIM the day before surgery. Blood samples for DIM levels were collected pre-treatment and just prior to surgery. DIM concentration was measured in the plasma and in prostate tissue specimens using a validated high-performance liquid chromatographic method with tandem mass spectrometric detection. AR was evaluated by immunohistochemistry (IHC). Results: 36 patients were treated at 2 institutions. The 26 evaluable patients had median age of 60 years (range 41 - 76), 14 were Caucasian, and 10 were African American. Reasons for inevaluability included change in surgery date (n=4), inadequate BR-DIM treatment (2), withdrawal from study (2), canceled surgery (1), and other (1). Toxicity was minimal; only two patients with grade 3 headache. Post dosing, DIM was found at a mean trough level of 10.2 ng/ml (range, 0.5 to 24.7) and 12.3 ng/gm of tissue (range, 0.0 to 26.8) in plasma and in prostate tissue, respectively. PSA levels had a slightly downward trend after BR-DIM treatment. In 18 of 20 evaluable PCa specimens, IHC showed nuclear exclusion of AR. Conclusions: BR-DIM was well tolerated, and DIM was detected in both plasma and prostate tissues at ~12 h post dosing. Nuclear exclusion of AR was found in 90% of PCa specimens post BR-DIM dosing, suggesting in vivo inactivation of AR activity. PSA levels were slightly reduced overall. Accrual is ongoing and nearly complete. Additional studies with BR-DIM in PCa are warranted.

scholarly journals Synthesis of Human Bone Morphogenetic Protein-2 (hBMP-2) in E. coli Periplasmic Space: Its Characterization and Preclinical Testing

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3525
Author(s):  
João E. Oliveira ◽  
Miriam F. Suzuki ◽  
Renata Damiani ◽  
Eliana R. Lima ◽  
Kleicy C. Amaral ◽  
...  
Keyword(s):  
High Performance ◽  
Osmotic Shock ◽  
Cytoplasmic Inclusion ◽  
Reversed Phase ◽  
C2c12 Cells ◽  
Size Exclusion ◽  
Bone Morphogenetic Protein 2 ◽  
E Coli

Human BMP-2, a homodimeric protein that belongs to the TGF- β family, is a recognized osteoinductor due to its capacity of inducing bone regeneration and ectopic bone formation. The administration of its recombinant form is an alternative to autologous bone grafting. A variety of E. coli-derived hBMP-2 has been synthesized through refolding of cytoplasmic inclusion bodies. The present work reports the synthesis, purification, and characterization of periplasmic hBMP-2, obtained directly in its correctly folded and authentic form, i.e., without the initial methionine typical of the cytoplasmic product that can induce undesired immunoreactivity. A bacterial expression vector was constructed including the DsbA signal peptide and the cDNA of hBMP-2. The periplasmic fluid was extracted by osmotic shock and analyzed via SDS-PAGE, Western blotting, and reversed-phase high-performance liquid chromatography (RP-HPLC). The purification was carried out by heparin affinity chromatography, followed by high-performance size-exclusion chromatography (HPSEC). HPSEC was used for qualitative and quantitative analysis of the final product, which showed >95% purity. The classical in vitro bioassay based on the induction of alkaline phosphatase activity in myoblastic murine C2C12 cells and the in vivo bioassay consisting of treating calvarial critical-size defects in rats confirmed its bioactivity, which matched the analogous literature data for hBMP-2.

scholarly journals Validation and Application of a New Reversed Phase HPLC Method forIn VitroDissolution Studies of Rabeprazole Sodium in Delayed-Release Tablets

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Md. Saddam Nawaz
Keyword(s):  
High Performance ◽  
Method Development ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Hplc Method ◽  
Array Detector ◽  
Relative Standard ◽  
Quality Control Tool ◽  
Rabeprazole Sodium

The purpose of this study was to develop and validate a new reversed phase high performance liquid chromatographic (RP-HPLC) method to quantifyin vitrodissolution assay of rabeprazole sodium in pharmaceutical tablet dosage form. Method development was performed on C 18,100×4.6 mm ID, and 10 μm particle size column, and injection volume was 20 μL using a diode array detector (DAD) to monitor the detection at 280 nm. The mobile phase consisted of buffer: acetonitrile at a ratio of 60 : 40 (v/v), and the flow rate was maintained at 1.0 mL/min. The method was validated in terms of suitability, linearity, specificity, accuracy, precision, stability, and sensitivity. Linearity was observed over the range of concentration 0.05–12.0 μg/mL, and the correlation coefficient was found excellent >0.999. The method was specific with respect to rabeprazole sodium, and the peak purity was found 99.99%. The method was precise and had relative standard deviations (RSD) less than 2%. Accuracy was found in the range of 99.9 to 101.9%. The method was robust in different variable conditions and reproducible. This proposed fast, reliable, cost-effective method can be used as quality control tool for the estimation of rabeprazole sodium in routine dissolution test analysis.

Xenopus laevis oocytes, eggs and tadpoles contain immunoactive insulin

1994 ◽  
Vol 141 (1) ◽  
pp. 123-129 ◽  
Author(s):  
F de Pablo ◽  
R Dashner ◽  
A R Shuldiner ◽  
J Roth
Keyword(s):  
Xenopus Laevis ◽  
High Performance ◽  
Immunoblot Analysis ◽  
Reversed Phase ◽  
Xenopus Laevis Oocytes ◽  
Maternal Origin ◽  
Insulin Mrna ◽  
Low Levels

Abstract Insulin is a multifunctional polypeptide hormone that regulates metabolic processes and promotes mitogenesis and differentiation in vitro in the cells and tissues of several species. Its role in vivo during embryogenesis is still poorly understood. We have previously found insulin mRNA in mature Xenopus laevis oocytes and in embryos during neurulation (before organogenesis of the pancreas takes place). We have now measured insulin immunoactivity in mature oocytes, unfertilized eggs and day-2 tadpoles. Using reversed phase high performance liquid chromatography, we found low levels of insulin in extracts of oocytes (stage VI). Both Xenopus insulin I and II were detected in unfertilized eggs. The day-2 tadpoles (stages 31–33) also contained immunoactive insulin, and in swimming tadpoles (stage 46) a few clusters of cells containing insulin immunoactivity could be identified by indirect immunofluorescence. Immunoblot analysis was relatively insensitive, detecting insulin only in the adult Xenopus pancreas. In summary, insulin (from maternal origin and embryonic expression) appears to be present early enough in Xenopus laevis to influence developmental processes such as neurulation. Journal of Endocrinology (1994) 141, 123–129

scholarly journals VALIDATION OF A BIOANALYTICAL REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR THE QUANTITATION OF NYSTATIN IN AN ANIMAL MODEL AFTER INTRANASAL IN SITU GEL ADMINISTRATION

2020 ◽  
pp. 180-184
Author(s):  
KAZI MARZUKA ◽  
DEHGHAN MOHAMED HASSAN
Keyword(s):  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Rat Plasma ◽  
Reversed Phase ◽  
Reverse Phase ◽  
Limit Of Quantification ◽  
Elution Time ◽  
Liquid Chromatographic

Objective: The aim of the study was to develop and validate a bioanalytical reverse-phase high-performance liquid chromatographic (HPLC) method for the estimation of nystatin in rat plasma after intranasal administration. Methods: The reversed-phase HPLC system was equipped with a Luna C18 column, the mobile system comprised of methanol, water, and dimethylformamide (55:30:15) and the flow rate was set at 0.9 ml/min. Results: The elution time for nystatin was 4.096±0.025 min. The calibration curves constructed in rat plasma were linear from 0.25 to 50 μg/ml. The lower limit of quantification (LOQ) was found to be 0.25 μg/ml. The standards for accuracy and precision of the intra- and inter-day variation studies were in the acceptable ranges as per the FDA guidelines. Conclusion: The LOQ value determined by the proposed method was noted to be satisfactory for inspecting the plasma pharmacokinetics of nystatin in rats’ post-administration of a nasal in situ gelling liquid crystalline precursor formulation in an in vivo study.

scholarly journals DETERMINATION OF 5H-BENZO[2,3][1,4]OXAZEPINO[5,6-B]INDOLES IN RAT PLASMA BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC-ULTRAVIOLET METHOD: APPLICATION TO PHARMACOKINETIC STUDIES

2017 ◽  
Vol 10 (12) ◽  
pp. 425 ◽  
Author(s):  
Ashok K Singh ◽  
Vinit Raj ◽  
Amit Rai ◽  
Amit K Keshari ◽  
Pranesh Kumar ◽  
...  
Keyword(s):  
High Performance ◽  
Gradient Elution ◽  
High Performance Liquid Chromatographic ◽  
Rat Plasma ◽  
Reversed Phase ◽  
Pharmacokinetic Studies ◽  
Relative Standard ◽  
Liquid Chromatographic

Objective: Recently, we reported newly synthesized 5H-benzo[2,3][1,4]oxazepino[5,6-b]indole) derivatives and proved their cytotoxicity against hepatocellular carcinoma specific Hep-G2 cell lines. We attempted herein to describe a reversed-phase high-performance liquid chromatographic method for the determination of three most active compounds 6a, 10a, and 15a in rat plasma to predict their pharmacokinetics parameters before in vivo study.Methods: A rapid and sensitive reversed-phase high-performance liquid chromatographic was employed for the determination of 6a, 10a, and 15a in rat plasma. Each compound was separated by a gradient elution of acetonitrile and water with 1 mL/min flow rate. The detector was set at 270, 285, and 275 nm for 6a, 10a, and 15a and the recorded elution times were 2.00, 2.87, and 1.88 min, respectively.Results: The calibration curve was linear with R2 of 0.938, 0.875, and 0.923 over the concentration range of 0.1–50 μg/mL. The inter- and intra-day variations of the assay were lower than 12.26%; the average recovery of 6a, 10a, and 15a was 97.31, 92.56, and 95.23 % with relative standard deviation of 2.12%, 3.25%, and 2.28%, respectively. The Cmax and Tmax were ~ 46.34, 18.56, and 25.65 μg/mL and 2.0, 4.0, and 4.0 h for 6a, 10a, and 15a, respectively, which indicate a robust method of detection in the present experiment.Conclusion: The study suggests that all of the three compounds have a lower rate of absorption, higher volume of distribution, and lower clearance rate, indicating good therapeutic response for in vivo activity. 

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