QUANTITATIVE ASSESSMENT OF THE PRINCIPAL STEROIDS FORMED BY HUMAN PLACENTAS PERFUSED WITH LABELLED CHOLESTEROL IN VITRO

1973 ◽  
Vol 72 (1) ◽  
pp. 127-136 ◽  
Author(s):  
G. Telegdy ◽  
M. Robin ◽  
E. Diczfalusy
Keyword(s):  
Human Placenta ◽  
Quantitative Assessment ◽  
Internal Standards ◽  
Carbon 14 ◽  
The Individual

ABSTRACT Three midgestation and three term placentas were perfused in vitro each with 5 mCi of [7α-3H] cholesterol. Perfusions were carried out at 37°C for 120 minutes, using diluted blood oxygenated with an air + CO2 mixture (23.1% O2 + 3.6% CO2). Radiochemically homogenous pregnenolone, progesterone, dehydroepiandrosterone and 17β-oestradiol were isolated from each placenta and each perfusate. Carbon-14 labelled internal standards were used in order to assess procedural losses. On an average 0.01 per cent of the perfused cholesterol was converted to the four steroids studied by midgestation placentas and 0.015 per cent by term placentas. Approximately equal amounts of the individual steroids were recovered from the placentas and from the perfusates. The amounts of pregnenolone + progesterone isolated exceeded those of dehydroepiandrosterone + 17β-oestradiol by a factor of 7 to 8. It is suggested that the human placenta is capable of converting circulating cholesterol not only to pregnenolone and progesterone, but also to dehydroepiandrosterone and 17β-oestradiol.

QUANTITATIVE ASSESSMENT OF THE DE NOVO STEROL AND STEROID SYNTHESIS IN THE HUMAN FOETO-PLACENTAL UNIT

1971 ◽  
Vol 66 (4) ◽  
pp. 666-678 ◽  
Author(s):  
D. F. Archer ◽  
R. S. Mathur ◽  
N. Wiqvist ◽  
E. Diczfalusy
Keyword(s):  
Sodium Acetate ◽  
Quantitative Assessment ◽  
De Novo ◽  
Isolation Procedure ◽  
Predominant Formation ◽  
Human Foetus ◽  
Steroid Synthesis ◽  
Internal Standards ◽  
Carbon 14 ◽  
Hydroxy Progesterone

ABSTRACT The quantitative significance of the de novo synthesis of Δ5- and Δ4-steroids was assessed in the midgestation human foetus. Two midgestation foetuses and two complete foeto-placental units were perfused at 36°C for 120 minutes with 3.0 and 6.0 mCi of [14C] sodium acetate, respectively, and radiochemically homogeneous carbon-14 labelled steroids were isolated from the adrenals, livers and perfusates. Losses throughout the isolation procedure were monitored in all tissues of all experiments by the recovery of tritium labelled internal standards. In the four experiments, a total of 18.0 mCi of [14C] sodium acetate was perfused and a total of 0.17 μCi of pregnenolone, 2.65 μCi of pregnenolone sulphate, 1.07 μCi of dehydroepiandrosterone and 8.64 μCi of dehydroepiandrosterone sulphate were isolated. A total of 12.53 μCi were isolated in the form of these four compounds; 10.13 μCi of this was present in the perfusates. In all, 0.07% of the administered [14C]-sodium acetate was converted to the four Δ5-steroids studied and 0.048% to dehydroepiandrosterone sulphate. Progesterone was isolated from the perfusates and liver extracts, and 17α-hydroxy-progesterone, androstenedione and testosterone from the perfusates. In the four experiments, a total of 0.045 μCi of carbon-14 labelled Δ4-steroids were isolated, predominantly from the perfusates. In all, less than 0.0003% of the perfused [14C] sodium acetate was converted to Δ4-steroids. Thus, almost 300 times more Δ5- than Δ4-steroids were formed. It is concluded that the extensive de novo steroidogenetic processes taking place in the midgestation human foetus lead to the predominant formation of Δ5-steroid sulphates.

IN VITRO STEROL AND STEROID SYNTHESIS BY OVARIES OF HYPOPHYSECTOMIZED IMMATURE RATS TREATED WITH HUMAN GONADOTROPHINS

1973 ◽  
Vol 73 (2) ◽  
pp. 321-334 ◽  
Author(s):  
R. Uma Bai ◽  
K. Bischoff ◽  
J. C. Macome ◽  
E. Diczfalusy
Keyword(s):  
Cholesteryl Esters ◽  
Cholesterol Esters ◽  
Steroid Synthesis ◽  
Internal Standards ◽  
Carbon 14 ◽  
Homogeneous Form ◽  
Immature Rats ◽  
Urinary Fsh

ABSTRACT The in vitro conversion of sodium [ 1,2- 14C] acetate into sterols and steroids was studied following the incubation of groups of ovaries from hypophysectomized immature rats with human gonadotrophins. Tritium labelled internal standards were used to monitor procedural losses and sterols and steroids were isolated in a radiochemically homogeneous form. The gonadotrophin preparations studied included a highly purified human urinary LH preparation (with no detectable FSH activity) as well as human urinary FSH and HCG preparations in which the LH and FSH-like activity, respectively, was selectively neutralized by the addition of appropriate antigonadotrophic sera. The ovaries of saline-treated control animals converted considerable quantities of labelled acetate into cholesterol and small amounts into cholesterol esters. Little, if any, acetate was converted by these ovaries into steroids. The injection of a human urinary LH-free FSH preparation or of a highly purified human urinary LH preparation resulted in a marked increase in the incorporation of labelled acetate into cholesterol, which was accompanied by a slightly increased incorporation into cholesteryl esters. Small amounts of carbon-14 labelled androstenedione and testosterone were also isolated but no pregnenolone or progesterone was found.

Schmerztherapie nach thoraxchirurgischen Eingriffen

2018 ◽  
Vol 12 (02) ◽  
pp. 155-165
Author(s):  
Holger Hendrix ◽  
Vladimir Kamlak ◽  
Georgi Prisadov ◽  
Katrin Welcker
Keyword(s):  
Thoracic Surgery ◽  
Pain Therapy ◽  
Pain Treatment ◽  
Internal Standards ◽  
Pain Experience ◽  
Thoracic Surgical Procedures ◽  
Therapeutic Algorithm ◽  
Alternative Procedures ◽  
The Individual ◽  
Documentation Form

The treatment of pain after thoracic surgery is a challenge and takes place in the individual clinics mostly according to clinic internal standards. It exists no currently valid S3 guideline for the treatment of acute perioperative and posttraumatic pain. For an effective pain treatment as well individual pain experience as the pain intensity of the various thoracic surgical procedures must be considered. Regular pain assessment with appropriate methods and their documentation form the basis for adequate and adapted pain therapy.There are a number of different pain therapy methods, non-medicamentous and drug-based methods, whose effectiveness is described in the literature partially different. For the treatment of acute postoperative pain after thoracic surgery, mainly drug-related procedures are used, except for physiotherapy as a non-medicamentous method. Increasingly, alternative procedures for the peridural catheter as a therapeutic gold standard in the treatment of pain after thoracic surgery are used. Their application can be integrated into a therapeutic algorithm.

scholarly journals In VitroAnalyses of the Effects of Heparin and Parabens on Candida albicans Biofilms and Planktonic Cells

2011 ◽  
Vol 56 (1) ◽  
pp. 148-153 ◽  
Author(s):  
Marisa H. Miceli ◽  
Stella M. Bernardo ◽  
T. S. Neil Ku ◽  
Carla Walraven ◽  
Samuel A. Lee
Keyword(s):  
Candida Albicans ◽  
Metabolic Activity ◽  
Biofilm Formation ◽  
High Dose ◽  
Dependent Manner ◽  
Planktonic Cells ◽  
Content Type ◽  
Heparin Sodium ◽  
The Individual

ABSTRACTInfections and thromboses are the most common complications associated with central venous catheters. Suggested strategies for prevention and management of these complications include the use of heparin-coated catheters, heparin locks, and antimicrobial lock therapy. However, the effects of heparin onCandida albicansbiofilms and planktonic cells have not been previously studied. Therefore, we sought to determine thein vitroeffect of a heparin sodium preparation (HP) on biofilms and planktonic cells ofC. albicans. Because HP contains two preservatives, methyl paraben (MP) and propyl paraben (PP), these compounds and heparin sodium without preservatives (Pure-H) were also tested individually. The metabolic activity of the mature biofilm after treatment was assessed using XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction and microscopy. Pure-H, MP, and PP caused up to 75, 85, and 60% reductions of metabolic activity of the mature preformedC. albicansbiofilms, respectively. Maximal efficacy against the mature biofilm was observed with HP (up to 90%) compared to the individual compounds (P< 0.0001). Pure-H, MP, and PP each inhibitedC. albicansbiofilm formation up to 90%. A complete inhibition of biofilm formation was observed with HP at 5,000 U/ml and higher. When tested against planktonic cells, each compound inhibited growth in a dose-dependent manner. These data indicated that HP, MP, PP, and Pure-H havein vitroantifungal activity againstC. albicansmature biofilms, formation of biofilms, and planktonic cells. Investigation of high-dose heparin-based strategies (e.g., heparin locks) in combination with traditional antifungal agents for the treatment and/or prevention ofC. albicansbiofilms is warranted.

scholarly journals Combined Luteolin and Indole-3-Carbinol Synergistically Constrains ERα-Positive Breast Cancer by Dual Inhibiting Estrogen Receptor Alpha and Cyclin-Dependent Kinase 4/6 Pathway in Cultured Cells and Xenograft Mice

Cancers ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 2116
Author(s):  
Xiaoyong Wang ◽  
Lijuan Zhang ◽  
Qi Dai ◽  
Hongzong Si ◽  
Longyun Zhang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cell ◽  
Breast Cancer Cells ◽  
Cultured Cells ◽  
Inhibitory Effect ◽  
Cyclin Dependent Kinase ◽  
Xenograft Mice ◽  
The Individual

The high concentrations of individual phytochemicals in vitro studies cannot be physiologically achieved in humans. Our solution for this concentration gap between in vitro and human studies is to combine two or more phytochemicals. We screened 12 phytochemicals by pairwise combining two compounds at a low level to select combinations exerting the synergistic inhibitory effect of breast cancer cell proliferation. A novel combination of luteolin at 30 μM (LUT30) and indole-3-carbinol 40 μM (I3C40) identified that this combination (L30I40) synergistically constrains ERα+ breast cancer cell (MCF7 and T47D) proliferation only, but not triple-negative breast cancer cells. At the same time, the individual LUT30 and I3C40 do not have this anti-proliferative effect in ERα+ breast cancer cells. Moreover, this combination L30I40 does not have toxicity on endothelial cells compared to the current commercial drugs. Similarly, the combination of LUT and I3C (LUT10 mg + I3C10 mg/kg/day) (IP injection) synergistically suppresses tumor growth in MCF7 cells-derived xenograft mice, but the individual LUT (10 mg/kg/day) and I3C (20 mg/kg/day) do not show an inhibitory effect. This combination synergistically downregulates two major therapeutic targets ERα and cyclin dependent kinase (CDK) 4/6/retinoblastoma (Rb) pathway, both in cultured cells and xenograft tumors. These results provide a solid foundation that a combination of LUT and I3C may be a practical approach to treat ERα+ breast cancer cells after clinical trials.

scholarly journals Antiviral Activity of the PropylamylatinTM Formula against the Novel Coronavirus SARS-CoV-2 In Vitro Using Direct Injection and Gas Assays in Virus Suspensions

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 415
Author(s):  
Ashley N. Brown ◽  
Gary Strobel ◽  
Kaley C. Hanrahan ◽  
Joe Sears
Keyword(s):  
Propionic Acid ◽  
Infectious Virus ◽  
Direct Injection ◽  
Plaque Assay ◽  
Antiviral Effect ◽  
Dependent Manner ◽  
Global Public Health ◽  
Novel Coronavirus ◽  
The Individual

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of novel coronavirus disease 2019 (COVID-19), has become a severe threat to global public health. There are currently no antiviral therapies approved for the treatment or prevention of mild to moderate COVID-19 as remdesivir is only approved for severe COVID-19 cases. Here, we evaluated the antiviral potential of a Propylamylatin formula, which is a mixture of propionic acid and isoamyl hexanoates. The Propylamylatin formula was investigated in gaseous and liquid phases against 1 mL viral suspensions containing 105 PFU of SARS-CoV-2. Viral suspensions were sampled at various times post-exposure and infectious virus was quantified by plaque assay on Vero E6 cells. Propylamylatin formula vapors were effective at inactivating infectious SARS-CoV-2 to undetectable levels at room temperature and body temperature, but the decline in virus was substantially faster at the higher temperature (15 min versus 24 h). The direct injection of liquid Propylamylatin formula into viral suspensions also completely inactivated SARS-CoV-2 and the rapidity of inactivation occurred in an exposure dependent manner. The overall volume that resulted in 90% viral inactivation over the course of the direct injection experiment (EC90) was 4.28 µls. Further investigation revealed that the majority of the antiviral effect was attributed to the propionic acid which yielded an overall EC90 value of 11.50 µls whereas the isoamyl hexanoates provided at most a 10-fold reduction in infectious virus. The combination of propionic acid and isoamyl hexanoates was much more potent than the individual components alone, suggesting synergy between these components. These findings illustrate the therapeutic promise of the Propylamylatin formula as a potential treatment strategy for COVID-19 and future studies are warranted.

scholarly journals Preclinical Testing of Radiopharmaceuticals for the Detection and Characterization of Osteomyelitis: Experiences from a Porcine Model

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4221
Author(s):  
Aage Kristian Olsen Alstrup ◽  
Svend Borup Jensen ◽  
Ole Lerberg Nielsen ◽  
Lars Jødal ◽  
Pia Afzelius
Keyword(s):  
Animal Model ◽  
Animal Models ◽  
Porcine Model ◽  
Animal Experiments ◽  
Radioactive Tracers ◽  
The Individual ◽  
Selection Of

The development of new and better radioactive tracers capable of detecting and characterizing osteomyelitis is an ongoing process, mainly because available tracers lack selectivity towards osteomyelitis. An integrated part of developing new tracers is the performance of in vivo tests using appropriate animal models. The available animal models for osteomyelitis are also far from ideal. Therefore, developing improved animal osteomyelitis models is as important as developing new radioactive tracers. We recently published a review on radioactive tracers. In this review, we only present and discuss osteomyelitis models. Three ethical aspects (3R) are essential when exposing experimental animals to infections. Thus, we should perform experiments in vitro rather than in vivo (Replacement), use as few animals as possible (Reduction), and impose as little pain on the animal as possible (Refinement). The gain for humans should by far exceed the disadvantages for the individual experimental animal. To this end, the translational value of animal experiments is crucial. We therefore need a robust and well-characterized animal model to evaluate new osteomyelitis tracers to be sure that unpredicted variation in the animal model does not lead to a misinterpretation of the tracer behavior. In this review, we focus on how the development of radioactive tracers relies heavily on the selection of a reliable animal model, and we base the discussions on our own experience with a porcine model.

scholarly journals Proteome Profiling of PMJ2-R and Primary Peritoneal Macrophages

2021 ◽  
Vol 22 (12) ◽  
pp. 6323
Author(s):  
Alexander L. Rusanov ◽  
Peter M. Kozhin ◽  
Olga V. Tikhonova ◽  
Victor G. Zgoda ◽  
Dmitry S. Loginov ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Living Organism ◽  
Peritoneal Macrophages ◽  
In Vitro Models ◽  
Individual Characteristics ◽  
Essential Proteins ◽  
Comparative Proteomic Analysis ◽  
Immortalized Cell ◽  
The Individual

In vitro models are often used for studying macrophage functions, including the process of phagocytosis. The application of primary macrophages has limitations associated with the individual characteristics of animals, which can lead to insufficient standardization and higher variability of the obtained results. Immortalized cell lines do not have these disadvantages, but their responses to various signals can differ from those of the living organism. In the present study, a comparative proteomic analysis of immortalized PMJ2-R cell line and primary peritoneal macrophages isolated from C57BL/6 mice was performed. A total of 4005 proteins were identified, of which 797 were quantified. Obtained results indicate significant differences in the abundances of many proteins, including essential proteins associated with the process of phagocytosis, such as Elmo1, Gsn, Hspa8, Itgb1, Ncf2, Rac2, Rack1, Sirpa, Sod1, C3, and Msr1. These findings indicate that outcomes of studies utilizing PMJ2-R cells as a model of peritoneal macrophages should be carefully validated. All MS data are deposited in ProteomeXchange with the identifier PXD022133.

Effects of Maternal Nutrient Restriction During Gestation on LH Production in the Female Bovine Fetus

2021 ◽  
Vol 99 (Supplement_2) ◽  
pp. 25-26
Author(s):  
Sterling H Fahey ◽  
Sarah West ◽  
John M Long ◽  
Carey Satterfield ◽  
Rodolfo C Cardoso
Keyword(s):  
Protein Content ◽  
Fetal Development ◽  
Monozygotic Twins ◽  
Late Gestation ◽  
Nutrient Restriction ◽  
Western Blots ◽  
Physiological Processes ◽  
Individual Potential ◽  
The Individual

Abstract Gestational nutrient restriction causes epigenetic and phenotypic changes that affect multiple physiological processes in the offspring. Gonadotropes, the cells in the anterior pituitary that secrete luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are particularly sensitive to nutritional changes during fetal development. Our objective herein was to investigate the effects of gestational nutrient restriction on LH protein content and number of gonadotropes in the fetal bovine pituitary. We hypothesized that moderate nutrient restriction during mid to late gestation decreases pituitary LH production, which is associated with a reduced number of gonadotropes. Embryos were produced in vitro with X-bearing semen from a single sire then split to generate monozygotic twins. Each identical twin was transferred to a virgin dam yielding four sets of female twins. At gestational d 158, the dams were randomly assigned into two groups, one fed 100% NRC requirements (control) and the other fed 70% of NRC requirements (restricted) during the last trimester of gestation, ensuring each pair of twins had one twin in each group. At gestational d 265, the fetuses (n = 4/group) were euthanized by barbiturate overdose, and the pituitaries were collected. Western blots were performed using an ovine LH-specific antibody (Dr. A.F. Parlow, NIDDK). The total LH protein content in the pituitary tended to be decreased in the restricted fetuses compared to controls (P &lt; 0.10). However, immunohistochemistry analysis of the pituitary did not reveal any significant changes in the total number of LH-positive cells (control = 460±23 cells/0.5 mm2; restricted = 496±45 cells/0.5 mm2, P = 0.58). In conclusion, while maternal nutrient restriction during gestation resulted in a trend of reduced LH content in the fetal pituitary, immunohistological findings suggest that these changes are likely related to the individual potential of each gonadotrope to produce LH, rather than alterations in cell differentiation during fetal development.

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