SUBCELLULAR DISTRIBUTION OF TESTOSTERONE METABOLITES AFTER PERFUSION OF ISOLATED LIVERS OF GUINEA PIG

1969 ◽  
Vol 62 (1) ◽  
pp. 147-152 ◽  
Author(s):  
K. Demisch ◽  
P. Birkelbach ◽  
U. Ammedick ◽  
W. Staib
Keyword(s):  
Subcellular Distribution ◽  
Extraction Procedure ◽  
Free Testosterone ◽  
List Type ◽  
Differential Centrifugation ◽  
Significant Difference ◽  
Testosterone Metabolites ◽  
Cell Fractions ◽  
Layer Chromatography ◽  
Steroid Conjugates

ABSTRACT The isolated livers of four guinea pigs were perfused with 4-14C-testosterone. It was found, that 23.1 % of the initially added activity was retained in the liver. The subcellular distribution of this 14C-activity was investigated by differential centrifugation. After the extraction procedure, the 14C-activity of the liver cell fractions was divided into A activity extractable by ether (free steroids), B activity remaining in the water phase (steroid-conjugates) and C activity associated with the filter residues (protein bound steroids). The ether extractable metabolites were separated by paper and thin layer chromatography and characterized by their gas chromatographic behaviour on a SE 30 and XE 60 column as free steroids, steroid-acetates and steroid-trimethylsilylethers. It was shown that there was no significant difference between the percentage distribution of free testosterone metabolites in the liver and perfusate. The steroid-conjugates had a three times higher concentration in the liver than in the perfusate. The subcellular distribution of free steroids showed significant differences. Of particular interest were the different ratios of testosterone to androstenedione in the subcellular fractions.

SUBCELLULAR DISTRIBUTION AND ARTERIO-VENOUS DIFFERENCES OF SYNTHETIC 3H-β1–23-CORTICOTROPHIN-23-AMIDE-ACETATE IN THE ADRENAL OF RATS

1969 ◽  
Vol 60 (3) ◽  
pp. 463-472 ◽  
Author(s):  
O. A. Müller ◽  
P. C. Scriba
Keyword(s):  
Single Dose ◽  
Cell Membrane ◽  
Thoracic Aorta ◽  
Subcellular Distribution ◽  
Mechanisms Of Action ◽  
Outer Cell ◽  
List Type ◽  
Differential Centrifugation ◽  
Outer Cell Membrane ◽  
Hypophysectomized Rats

ABSTRACT Synthetic 3H-β1–23-corticotrophin-23-amide-acetate (3H-ACTH) was given as a single dose by means of a 95 s infusion into the thoracic aorta of hypophysectomized rats. Distribution of radioactivity was determined in adrenal subcellular fractions prepared by homogenization and differential centrifugation: 3, 12, 40 and 120 min after the start of the 3H-ACTH infusion. Radioactivity in nuclear fractions, given as a % of total adrenal CPM rose from 27 % (3 min) to 37 % (40 and 120 min, P < 0.0025). These results may indicate ACTH penetration through the outer cell membrane and interaction with adrenal nuclei. Microsomal radioactivity (18 %) at 3 min after the start of 3H-ACTH infusion appeared to be in excess of the contribution of this fraction to the total adrenal weight and decreased significantly with time (120 min: 3 %, P < 0.0005). These observations are discussed with regard to mechanisms of action of ACTH. When synthetic 3H-β1–23-corticotrophin-23-amide-acetate was infused into the thoracic aorta of rats, the differences in the radioactivity of the plasma from the aorta and from the adrenal vein were significantly higher (P < 0.0125) in hypophysectomized than in sham-operated rats, presumably indicating in the latter the saturation by endogenous ACTH of adrenal receptors for ACTH.

scholarly journals Comparison between Pressurized Liquid Extraction and Conventional Soxhlet Extraction for Rosemary Antioxidants, Yield, Composition, and Environmental Footprint

Foods ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 584 ◽  
Author(s):  
Mathilde Hirondart ◽  
Natacha Rombaut ◽  
Anne Sylvie Fabiano-Tixier ◽  
Antoine Bily ◽  
Farid Chemat
Keyword(s):  
Extraction Procedure ◽  
Soxhlet Extraction ◽  
Liquid Extraction ◽  
Raw Material ◽  
Pressurized Liquid Extraction ◽  
Carnosic Acid ◽  
Environmental Footprint ◽  
Initial Composition ◽  
Significant Difference ◽  
Preparation Step

Nowadays, “green analytical chemistry” challenges are to develop techniques which reduce the environmental impact not only in term of analysis but also in the sample preparation step. Within this objective, pressurized liquid extraction (PLE) was investigated to determine the initial composition of key antioxidants contained in rosemary leaves: Rosmarinic acid (RA), carnosic acid (CA), and carnosol (CO). An experimental design was applied to identify an optimized PLE set of extraction parameters: A temperature of 183 °C, a pressure of 130 bar, and an extraction duration of 3 min enabled recovering rosemary antioxidants. PLE was further compared to conventional Soxhlet extraction (CSE) in term of global processing time, energy used, solvent recovery, raw material used, accuracy, reproducibility, and robustness to extract quantitatively RA, CA, and CO from rosemary leaves. A statistical comparison of the two extraction procedure (PLE and CSE) was achieved and showed no significant difference between the two procedures in terms of RA, CA, and CO extraction. To complete the study showing that the use of PLE is an advantageous alternative to CSE, the eco-footprint of the PLE process was evaluated. Results demonstrate that it is a rapid, clean, and environmentally friendly extraction technique.

scholarly journals Triglycerides, independent of Ferriman Gallwey Score, is a main determinant of free testosterone index in PCOS

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 94
Author(s):  
Andon Hestiantoro ◽  
Putri Deva Karimah ◽  
Amalia Shadrina ◽  
Budi Wiweko ◽  
R. Muharam ◽  
...  
Keyword(s):  
Free Testosterone ◽  
Cross Sectional Study ◽  
Pcos Patient ◽  
Reproductive Age ◽  
Bivariate Analysis ◽  
Sample Collection ◽  
Cross Sectional ◽  
Significant Difference ◽  
Shbg Level ◽  
Metabolic Dysfunctions

Background: Polycystic Ovarian Syndrome (PCOS) is the most common endocrinopathy in women of reproductive age, affecting 5-20% of women worldwide. Hyperandrogenism, as the primary characteristic of PCOS, is not always present in every patient. The hyperandrogenic phenotype of PCOS patients is influenced by both hormonal and metabolic dysfunctions. Therefore, this study aims to determine the correlation between hormone profile, lipid profile, and clinical profile with free testosterone index in subjects with PCOS. Methods: This prospective cross-sectional study was conducted in the Dr. Cipto Mangunkusumo General Hospital between July 2014 and December 2016. The study involved 76 women with PCOS, who were classified into 2 subgroups: 39 subjects in the hyperandrogenism group and 37 subjects in the non-hyperandrogenism group. Each subject underwent physical examination, blood sample collection, and USG examination. Bivariate analysis was done using independent t-tests and Mann Whitney U-tests, while multivariate analysis was done using logistic regression. Results: Triglyceride and testosterone level showed weak (r = 0.232, p = 0.044) and moderate (r = 0.460, p ¡ 0.001) positive correlation with FTI, while SHBG level showed moderate negative correlation (r = -0.483, p ¡ 0.001). Triglyceride was also found to be determinant of hyperandrogenism condition in PCOS patient (OR 0.02, 95% CI 0.00–0.04, p = 0.013). However, there was no significant difference observed between FGS and hyperandrogenism (p = 0.43). Conclusions: Triglycerides, testosterone, and SHBG were associated with hyperandrogenism in PCOS patients, while FGS showed no such association.

An Automatic Handheld Device for Firmness Measurement of Fruit with Edible Peel Based on Resistance Sensing: A Case Study of Peaches

2021 ◽  
Vol 64 (6) ◽  
pp. 1895-1905
Author(s):  
Dachen Wang ◽  
Zhe Feng ◽  
Di Cui
Keyword(s):  
3D Models ◽  
Initial Slope ◽  
List Type ◽  
Fruit Firmness ◽  
Important Indicator ◽  
Handheld Device ◽  
Relative Standard ◽  
Significant Difference ◽  
Measurement Results

HighlightsA handheld device was developed for automatic on-site measurement of fruit firmness.The prototype device achieved better performance than a typical commercial penetrometer.Two commonly used firmness-related parameters could be predicted based on the prototype readings.The distribution and changes of firmness could be visualized based on 3D models of the prototype readings.Abstract. Firmness is an important indicator of fruit quality. The devices for on-site fruit firmness measurement need to be portable, low-cost, lightweight, reliable, and flexible. Existing portable devices can provide firmness-related parameters, but their measurement results cannot be converted to each other. Their measurement results are also susceptible to operator error due to the manual measurement process. To solve this problem, a prototype handheld device was developed based on sensing fruit resistance to achieve automatic on-site firmness measurement of fruit with edible peel and provide commonly used firmness-related parameters. The prototype’s precision was verified by analyzing the repeatability and reproducibility of the measurement results with four Shore hardness blocks. The relative standard deviations (RSDs) of the results obtained by the same operator were less than 0.61%, and there was no significant difference among the results obtained by different operators (p &lt; 0.05), which indicated that the prototype could provide reliable measurement results. A case study of peaches was performed, and the results showed that (1) the prototype had better performance than a commercial penetrometer, (2) two commonly used firmness-related parameters could be predicted based on regression models between a texture analyzer and the prototype readings (R2P = 0.908 and RMSEP = 4.191 N for maximum force; R2P = 0.923 and RMSEP = 1.613 N mm-1 for initial slope), (3) changes in the prototype readings for peaches during growth corresponded with the growth characteristics, and (4) the distribution and changes of peach firmness over time could be visualized based on 3D models of the prototype readings. Keywords: Automatic, Fruit firmness, Handheld device, High precision, Peach.

Abstract 2244: A Novel Micropace-Ablate Protocol Eliminates Coagulum Formation in Radiofrequency Ablation

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Bernard Lim ◽  
K.l. Venkatachalam ◽  
Susan Johnson ◽  
Arshad Jahangir ◽  
Samuel Asirvatham
Keyword(s):  
Surface Coverage ◽  
Negative Charge ◽  
Fibrin Clot ◽  
List Type ◽  
Rf Ablation ◽  
Persistent Problem ◽  
Significant Difference ◽  
Catheter Tip ◽  
Delivery Unit ◽  
Rf Energy

Radiofrequency (RF) ablation for cardiac arrythmias is an established curative therapy. However, catheter-tip coagulum formation leading to thromboembolic stroke is a persistent problem despite heparinization. Recent studies demonstrated an up to 70% incidence of thrombus formation on the catheter tip during ablation and a 10 % incidence of thromboembolic events. Catheter tip thrombus can impede the delivery of RF energy to the tissue, resulting in reduced efficacy and longer procedure times. Here we report on a novel method to prevent coagulum formation on ablation catheters by delivering a negative charge to the catheter tip to repel negatively charged fibrinogen molecules during RF ablation. A novel circuit was built with a charge delivery unit using a 9 voltage battery to deliver a fixed offset direct current charge, placed in parallel to the RF delivery unit for negative charge placement during RF delivery. In in-vivo canine experiments (n=50) standard 6/4, 8/5 ablation catheters and catheters with abraded tips were advanced from right femoral veins into atria and ventricles under intra-cardiac echo (ICE) and biplane fluoroscopic guidance. The dogs received 1 of 2 therapies: no RF energy delivered with and without negative charge delivery, RF energy delivery with and without negative charge delivery. The presence of thrombus was identified with ICE. The catheter tips were examined also macroscopically with visual inspection and also with field emission scanning electron microscopy (FESEM). FESEM of the catheter tips showed surface coverage of fibrin clot of the catheter tip to be 90 % for non-charged catheters compared to 0 % (p< 0.01) in negatively-charged catheters. There was no significant difference in surface coverage of fibrin clot between negatively-charged catheters with the abraded tips (98.8%) and non-charged catheters with smooth tips (90.7 %, p=ns). Clot volume formed on the catheter tip decreased with increased amount of charge. Application of a negative charge did not affect the quality of the intracardiac electrograms or induce malignant ventricular arrhythmias. Negative charge delivery during RF ablation is feasible and safe and can eliminate coagulum formation, potentially reducing thromboembolic complications.

scholarly journals ζ-, ε-, and γ-Globin mRNA in Blood Samples and CD71+ Cell Fractions from Fetuses and from Pregnant and Nonpregnant Women, with Special Attention to Identification of Fetal Erythroblasts

2001 ◽  
Vol 47 (4) ◽  
pp. 645-653 ◽  
Author(s):  
Anne Mette Høgh ◽  
Thomas Vauvert F Hviid ◽  
Britta Christensen ◽  
Steen Sørensen ◽  
Rasmus D Larsen ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Rt Pcr ◽  
Globin Mrna ◽  
Isolation And Identification ◽  
Blood Samples ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Significant Difference ◽  
Fisher’S Exact Test ◽  
Cell Fractions

Abstract Background: Information about the appearance of γ-, ε-, and ζ-globin mRNAs in fetal erythroblasts during gestation and about the presence and amounts of these mRNAs in pregnant and nonpregnant women is important from the perspective of using these molecules as a marker of fetal erythroblasts. A specific marker is necessary for isolation and identification of fetal nucleated red blood cells from maternal blood samples for use in antenatal diagnosis of fetal genetic or chromosomal abnormalities. Methods: We used a very sensitive reverse transcription-PCR (RT-PCR) method, coamplification analysis of γ- and ε-globin cDNA, and quantitative analysis of γ-globin mRNA based on competitive RT-PCR to investigate these aspects. Results: All adult whole-blood samples were negative for ε- and ζ-globin mRNA. Analyses of CD71+ cell fractions showed that specimens from 19 of 20 nonpregnant and 10 of 14 pregnant women (at 9–13 weeks of gestation) were positive for γ-globin mRNA (Fisher’s exact test, P = 0.13), and those from 3 of 20 nonpregnant and 5 of 14 pregnant women were positive for ζ-globin mRNA (Fisher’s exact test, P = 0.23). No ε-globin mRNA was detected in CD71+ cell fractions from 1-mL blood samples from adults. CD71+ cell fractions from eight fetal blood samples (at 17–20 weeks of gestation) were positive for all three globin mRNAs. We found no statistically significant difference between the amounts of γ-globin mRNA in pregnant and nonpregnant women. Conclusions: This study indicates that ε-globin mRNA might function as a marker for fetal CD71+ cells early in pregnancy. Although γ-globin mRNA can be detected in CD71+ cell fractions from most adults, these transcripts also may be of use because of a marked difference between adult and fetal values.

scholarly journals Isolation of Chloramphenicol from Whole Eggs by Supercritical Fluid Extraction with In-Line Collection

1999 ◽  
Vol 82 (6) ◽  
pp. 1334-1339 ◽  
Author(s):  
John W Pensabene ◽  
Walter Fiddler ◽  
Dan J Donoghue
Keyword(s):  
Solvent Extraction ◽  
Supercritical Fluid Extraction ◽  
Supercritical Fluid ◽  
Extraction Method ◽  
Gas Flow ◽  
Extraction Procedure ◽  
The United States ◽  
Fluid Extraction ◽  
Solvent Extraction Method ◽  
Significant Difference

Abstract Egg consumption, at more than 65 billion per year in the United States, represents a potentially significant source of exposure to drug residues, particularly if the laying hens are treated with antimicrobial compounds or fed a diet containing medicated feed. Residues resulting from the use of chloramphenicol (CAP) is especially problematic if this compound is not used in accordance with national registration, e.g., for the control of Salmonella microorganisms in poultry. The most commonly used methods for the determination of CAP in biological samples require the use of large amounts of organic solvent. As a result, a less solvent intensive supercritical fluid extraction (SFE) method was developed for CAP in whole chicken eggs, and the results were compared with those for a solvent extraction procedure. In the SFE method, the egg sample is extracted with supercritical CO2 (without a modifier) at 10 000 psi (680 bar), 80°C, and an expanded gas flow rate of 3.0 L/min to a total volume of 150 L. The CAP is trapped in-line on a Florisil sorbent bed. The CAP is eluted post-SFE by using the liquid chromatographic mobile phase solvent (water-methanol), and determined on a C8 column with ultraviolet detection at 280 nm. Recovery from eggs fortified at the 10 ppb level (n = 6) was 81.2 ± 4.3%. To obtain eggs containing incurred CAP, hens were given a single daily dose of 75 mg CAP (orally by gelatin capsule) for 2 consecutive days, and the eggs were collected over a 12-day period. The mean value for “normally incurred” CAP in the eggs (n = 17) analyzed by SFE ranged from none detected to 174.5 ppb, with an overall mean of 60.5 ppb, compared with a mean of 60.4 ppb for the solvent extraction method. No significant difference in results was found between methods. However, the SFE method is more rapid, uses less solvent, and gives recoveries similar to those for the solvent extraction method, making it ideal for regulatory monitoring.

Improved one-dimensional thin-layer chromatography of phospholipids in amniotic fluid.

1982 ◽  
Vol 28 (1) ◽  
pp. 209-211 ◽  
Author(s):  
A A Pappas ◽  
R E Mullins ◽  
R H Gadsden
Keyword(s):  
Amniotic Fluid ◽  
Mobile Phase ◽  
Distress Syndrome ◽  
Extraction Procedure ◽  
Fetal Lung ◽  
One Dimensional ◽  
Cupric Acetate ◽  
Rf Values ◽  
Layer Chromatography ◽  
Lung Maturity

Abstract With this extraction procedure and chromatographic method, six amniotic fluid phospholipids [phosphatidylcholine (lecithin), sphingomyelin, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, and phosphatidylglycerol] are completely resolved in less than 70 min from the time the specimen is received. The mobile phase (chloroform/petroleum ether/methanol/acetic acid, 5/3/1.6/1, by vol) is used with commercially available 7.5 X 10 cm silica-gel plates. The phospholipids are made visible by immersing the plate in cupric acetate/phosphoric acid. Lecithin/sphingomyelin (L/S) ratios are determined densitometrically. Rf values are well reproducible for both samples and standards, as are L/S ratios, and the method is simple and inexpensive. The relation between L/S ratios (for 100 specimens of amniotic fluid measured by this method) and fetal status is consistent with other reports. We confirm phosphatidylglycerol to be a sensitive indicator of fetal lung maturity: when it was detectable in amniotic fluid, the newborn invariably showed no symptoms of the respiratory distress syndrome.

P06 A study to assess the effectiveness of vitamin d supplementation in paediatric cystic fibrosis patients

2018 ◽  
Vol 103 (2) ◽  
pp. e1.9-e1
Author(s):  
Christiansen Nanna ◽  
Ashraf Saleha
Keyword(s):  
Cystic Fibrosis ◽  
Vitamin D ◽  
Vitamin D Deficiency ◽  
Dose Range ◽  
Vitamin D Supplementation ◽  
High Dose ◽  
List Type ◽  
Current Guideline ◽  
Paediatric Patients ◽  
Significant Difference

AimsPatients with cystic fibrosis (CF) require supplementation of fat-soluble vitamins due to the effects of the disease on the pancreas and the resulting inability of absorb fat effectively.1The study aimed to assess the effectiveness of current of vitamin D supplementation to achieve adequate serum Vitamin D (25OHD) levels in paediatric CF patients.2 Secondly, this study assessed the effectiveness of ‘Stoss’ therapy (a high dose vitamin D therapy administered every three months) as an alternative to daily vitamin D supplementation for patients with known poor compliance.3MethodsVitamin D doses and serum 25OHD levels between January and December 2016 were reviewed for paediatric CF patients at a UK centre. Data was collected for 138 paediatric patients. The ‘clinical record summary’ system was used to extract the data which included age, hospital number, weight in 2015 and 2016, 25OHD levels from 2015 and 2016, vitamin D dose before each level and pancreatic status.Data was entered onto Statistical Package for the Social Sciences (SPSS) system for analysis. A paired T-test was conducted to ascertain if there was a significant difference in weekly/kg doses between patients that were sufficient (25OHD>50 nmol/L) and insufficient (25OHD<50 nmol/L).ResultsData was collected for a total of 138 patients. The data from only 70 patients was analysed when investigating the first objective, as all other patients did not have 25OHD levels available for both 2015 and 2016. A further five patients wereexcluded and analysed seperately due to receiving Stoss therapy. The weekly Vitamin D dose range was very wide for both years with 43% (n=40) of patients requiring additional vitamin D in addition to Aquadeks (CF multivitamin preparation). There was no significant difference in Vitamin D doses between patients with sufficient and insufficient 25OHD levels. This was thecase for both 2015 (p=0.432) and 2016 (p=0.192). The daily supplementation doses were successful at maintaining vitamin D sufficiency for 83% of patients in 2015 and 93% in 2016.Out of the 5 patients who received ‘Stoss’ therapy, 3 had an increase in 25OHD levels. However, only one of the patients had a significant increase leading to sufficient 25OHD levels. In 2 cases there was actually a 60%–68% decrease in 25OHD levels, which lead to these patients developing vitamin D deficiency.ConclusionThis study was useful in determining the effectiveness of current Vitamin D dosing. The results suggest that patients having insufficient 25OHD levels may not be due to an inadequacy of doses provided in the current guideline, as there was no significant difference in dose between patients with sufficient and insufficient 25OHD levels. Given the patient group, the difference could be attributable to a lack of compliance to daily therapies in the patients with insufficient 25OHD levels or even differences in individual responses to therapy.In this sample, ‘Stoss’ therapy is not effective in maintaining sufficient 25OHD levels. Although the data for this part of the study was very limited, it identifies a need to investigate the effectiveness of ‘Stoss’ therapy further.ReferencesFerguson JH, Chang AB. Vitamin D supplementation for cystic fibrosis. Cochrane Database of Syst Rev [Internet] 2014. http://onlinelibrary.wiley.com/ & doi:10.1002/14651858.CD007298.pub3/pdf [Available: 2017April 12].Green D, Carson K, Leonard A, et al. Current treatment recommendations for correcting vitamin D deficiency in paediatric patients with cystic fibrosis is inadequate. J Pediatr2008;4:554–559.Shepherd D, Belessis Y, Katz, et al. Single high-dose oral vitamin D3 (stoss) therapy: A solution to vitamin D deficiency in children with cystic fibrosis?J Cyst Fibros2013;2:177–182.

Simple Manual Procedure for Determination of Serum Triglycerides

1975 ◽  
Vol 21 (6) ◽  
pp. 768-770 ◽  
Author(s):  
Jose Mendez ◽  
Barry Franklin ◽  
Harry Gahagan
Keyword(s):  
Quality Control ◽  
Calibration Curve ◽  
Room Temperature ◽  
Extraction Procedure ◽  
Serum Triglycerides ◽  
Modified Method ◽  
Color Development ◽  
Significant Difference ◽  
Control Samples

Abstract We describe a modified method for determining serum triglycerides (triacylglycerols), which is based on the heptane extraction procedure of Gottfried and Rosenberg [Clin. Chem. 19, 1077 (1973)] with the stable saponification, oxidation, and color development reagents of Neri and Frings [Clin. Chem. 19, 1201 (1973)]. This modified method eliminates one heating step, reduces saponification time to 5 min, absorbances are read at room temperature, and the calibration curve is linear to 3.0 g/liter. A sample comparison between the proposed method and the automated Block and Jarrett [Am. J. Med. Technol. 35, 1 (1969)] procedure showed no significant difference (r = 0.98). The coefficient of variation (47 duplicate samples) for the modified method was 6.3%. Further validation was obtained from analysis of quality-control samples; the proposed method gave equivalent values.

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