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Influence of Progesterone on Orthopox Viruses in vitro and in vivo

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Abstract

The influence of progesterone on virus-host-interactions was investigated for orthopox viruses in vitro and in vivo. Virus induced cytopathogenic effects in Vero cells were markedly reduced in presence of the hormone. Dose dependent restriction of viral replication was evaluated by infectivity titration and immunological antigen determination. Progesterone concentrations of 5μg/ml tissue culture medium depressed the production of new viral particles nearly totally. The results obtained by the two different methods indicate that probably later stages of viral synthesis are affected when lower dosages of progesterone are used. The protective activity of progesterone against pathogenic actions of orthopox viruses in vivo was confirmed by using rabbits for intradermal infections. Es wurde der Einfluß von Progesteron auf die Virus-Wirt-Wechselbeziehung am Beispiel der Orthopocken-Viren in vitro und in vivo untersucht. Die virusinduzierten zytopathogenen Effekte in Vero-Zellkulturen wurden in Gegenwart des Hormons deutlich reduziert. Die Hemmung der Virus-Replikation in Abhängigkeit von der Dosis wurde mittels Infektionstitration sowie immunologischer Antigenbestimmung gemessen. Eine Progesteron-Konzentration von 5 μg/ml Zellkulturmedium unterdrückte die Bildung von Viruspartikeln nahezu vollständig. Die Ergebnisse der beiden unterschiedlichen Methoden lassen vermuten, daß durch niedrigere Progesteron-Konzentrationen spätere Stadien der Virussynthese gehemmt werden. Durch intradermale Infektion von Kaninchen konnte die protektive Wirkung von Progesteron gegen zytopathogene Mechanismen der Orthopox-Viren in vivo bestätigt werden.

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... Progesterone (S5) is a sex steroid hormone and the major progestogen in the body. It was found to inhibit orthopox virus in late stages of infection in Vero cells and in live rabbits (Pfahler, Reimann, and Munz 1987). Interestingly, VACV SalF7L (A44L) gene product encodes a protein with 31% identity to 3β-HSD, the enzyme that catalyses the synthesis of progesterone from pregnenolone, and its deletion led to attenuated virulence Smith 1992, Reading, Moore, and. ...
Thesis
Poxviruses are a family of large, double-stranded DNA viruses that infect a wide range of organisms, from insects to humans. They possess a complex structure and an intricate replication cycle that takes place entirely in the cytoplasm. Additionally, their genome is remarkably large for viruses, encoding over 200 proteins. Historically, one representative of the orthopox genera – variola virus, the causative agent of smallpox – has claimed hundreds of millions of human lives. Despite its eradication, smallpox, and some zoonotic poxviruses, are still considered as potential threats, and new means of combating them are in demand. To this end research has continued on vaccinia virus (VACV), the prototypic orthopox virus which was used as the smallpox vaccine. In this project a library of 1,280 FDA approved molecules was screened for cell-based VACV inhibitors using a high-throughput image-based screen, following the principles of drug repurposing. After confirmation of initial hits using secondary screens, 9 Early inhibitors, 3 Late inhibitors, and 7 Spread inhibitors were identified. Additionally, 14 cardiac glycosides were identified as Early inhibitors. Using focused assays, the specific phases of the virus replication cycle at which inhibition occurs were also identified. This project led to the discovery of new compounds that block VACV infection at different stages in cell culture. Additionally, the initial hits were tested on African swine fever virus and 5 potential inhibitors were identified. Collectively, these results have furthered our insights into host – pathogen interaction, while providing potential hits for testing in animal models and against other viruses.
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[4-14C]Progesterone was incubated with homogenate and mitochondrial, microsomal and soluble fraction preparations of healthy and inflamed gingiva from human subjects of both sexes. The subcellular preparations were supplemented with an NADPH-regenerating system and incubated for 2 h at pH 7.4 and 37 degrees C. The metabolites were identified by column, multiple TLC and radioautography and quantified with liquid scintillation counting. In inflamed tissue the metabolic activity was higher than in healthy gingiva. On the basis of the identified metabolites it can be concluded that the human gingiva of both sexes contains marked 3 alpha-, 3 beta- and 20 alpha-hydroxysteroid dehydrogenase, delta 4-5 alpha- and delta 4-5 beta-steroid hydrogenase activities, and less 20 beta-hydroxysteroid dehydrogenase activity.
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Yotis, William (Loyola University, Hines, Ill.), and Ronald Stanke. Bacteriostatic action of progesterone on staphylococci and other microorganisms. J. Bacteriol. 92 1285–1289. 1966.—Progesterone has been examined in vitro for antibacterial activity against 10 microorganisms. Turbidimetric and manometric techniques were used to assay the antibacterial activity of progesterone. The organisms tested consisted of Staphylococcus aureus, S. epidermidis, Gaffkya tetragena, Bacillus subtilis, Listeria monocytogenes, Candida albicans, Escherichia coli, Aerobacter aerogenes, Salmonella paratyphi, and Proteus vulgaris. Antibacterial action was shown by progesterone only against the gram-positive microorganisms when they were grown in tryptic soy broth containing 10 to 20 μg of progesterone per ml. Pregnenolone, 4-pregnen-20β-ol-3-one, and 5α-pregnane also possessed antistaphylococcal properties, whereas pregnanolone, pregnandione, 11α-hydroxyprogesterone, and 17α-hydroxyprogesterone did not. The bacteriostatic action of progesterone on staphylococci was exerted primarily during the first 8 hr of incubation, and it was reduced in the presence of oxygen. In the presence of 20 μg of progesterone per ml, there was significant reduction in the oxidation by resting staphylococcal suspensions or utilization by staphylococci of pyruvate as an energy source during growth.
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The yield of Aspergillus niger mycelium from a synthetic medium can be increased by the addition of microgram quantities of cholesterol, ergosterol, cholestanol, 7-dehydrocholesterol, stigmasterol, sitosterol, pregnenolone, and the vitamins D. The stimulation is not due to degradation to the acetate level. It is obtained only in highly aerated cultures. The rate of growth of Torula utilis was not increased. Both organisms were inhibited by desoxycorticosterone, testosterone, androstenedione, cortisone acetate, progesterone, and diethylstilbestrol. T. utilis was also inhibited by estradiol. A small decrease in progesterone inhibition of T. utilis was obtained by adding ergosterol, cholesterol, or pregnenolone. Of the compounds which have been adequately tested the order of stimulatory activity for A. niger is: ergosterol > cholesterol > stigmasterol > 7-dehydrocholesterol > cholestanol > pregnenolone. Progesterone was inhibitory at low concentrations but stimulatory at higher ones, while 17-hydroxyprogesterone was neither inhibitory nor stimulatory. Desoxycorticosterone and testosterone were inhibitory at all concentrations. Complete inhibition of the growth of the fungus was not obtained with any of the steroids. It is concluded that A. niger has a metabolic requirement for a steroid with a hydroxy group on carbon 3, a double bond in the 5–6 position, and a side chain similar to that in ergosterol or cholesterol and that this material is growth-limiting in the early stages of the cultures described.
Progesterone binding and inhibition of growth in Trichophyton mentagrophytes Immunsuppres-sion, sialic acid, and sialyltransferase of bovine serum as a function of progesterone concentra-tion
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