IN VIVO ANDROGEN BINDING IN RAT SKELETAL AND PERINEAL MUSCLES

ABSTRACT In vivo binding of [3H] testosterone2), [3H]5α-dihydrotestosterone and [3H]3α-androstanediol to cytosolic and nuclear fractions of LA/BC and thigh muscles has been studied in functionally hepatectomized castrated rats following a 1 h infusion of the labelled steroid. The identification of metabolites formed from each steroid has also been determined in tissue cytosols. In each experiment, ventral prostate was used as reference target tissue. After [3H] testosterone and [3H]5α-dihydrotestosterone perfusions, cytosolic binding could be demonstrated in a 8–10S peak on sucrose gradient with LA/BC and ventral prostate or in the macromolecular fraction after filtration through Sephadex G-25 with thigh muscles. In both types of muscles, [3H]5α-dihydrotestosterone binding represented only one tenth of [3H] testosterone binding. This behaviour seems to be related to the high rate of 5α-dihydrotestosterone metabolism observed in these tissues; testosterone, on the contrary, was not metabolized. After [3H]3α-androstanediol perfusion, cytosolic [3H] androgen binding in LA/BC and in thigh muscles was almost non-existent. In muscles [3H]3α-androstanediol remained essentially unconverted. In ventral prostate, with every hormone studied level of cytosolic binding was comparable. It was observed that in this tissue [3H] testosterone and [3H]3α-androstanediol were metabolized into [3H]5α-dihydrotestosterone. Androgen binding to 0.4 m KCl extracted nuclear proteins has been demonstrated in ventral prostate as a 3.5–4.5S binding peak on sucrose gradient and this with each steroid perfused. In LA/BC, only [3H] testosterone gave a well defined binding peak. In thigh muscles, levels of nuclear binding were too low to be determined. In summary, these results suggest that rat perineal and skeletal muscles possess cytosolic androgen binding proteins similar to those found in ventral prostate. However, it appears that steroid metabolism is quite different in ventral prostate and muscles with respect to presence of 5α-reductase activity and extent of conversion of 5α-dihydrotestosterone into androstanediols. These differences may explain why, in vivo, muscles bind testosterone instead of 5α-dihydrotestosterone as in ventral prostate.

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A model for studies on the response of the ventral prostate to oestrogens

Acta Endocrinologica ◽

10.1530/acta.0.0970125 ◽

1981 ◽

Vol 97 (1) ◽

pp. 125-136 ◽

Cited By ~ 10

Author(s):

J. J. Corrales ◽

P. A. Høisaeter ◽

N. Kadohama ◽

G. P. Murphy ◽

A. A. Sandberg

Keyword(s):

Body Weight ◽

Organ Culture ◽

Age Differences ◽

Oestrogen Receptor ◽

Direct Interaction ◽

Ventral Prostate ◽

Target Tissue ◽

The Mean ◽

In Vivo Effects

Abstract. The effects of oestrogen administration on the weight of ventral and dorsolateral prostates were studied in castrated rats of Wistar-Furth and Copenhagen strains. Direct effect of oestradiol (Oe2) on prostatic tissue was also investigated in organ culture. Oe2-treated animals received daily injections of 50 μg for 7 days. Control animals were treated with the vehicle only (peanut oil). In 4 month old Copenhagen rats the mean weight of the ventral prostates (42.4 ± 9.4 mg/100 g body weight) was significantly higher than that in the control animals (19.9 ± 4.6 mg/100 g body weight, P < 0.0001). No such differences were observed in older Copenhagen rats (9 months old) or Wistar-Furth rats (3 and 6 months old). Thus, this effect of Oe2 on ventral prostate seems both strain specific and age specific. Similar strain and age differences in the Oe2 effect were found in the dorsolateral prostate, but to a smaller extent. Direct interaction of Oe2 with target tissue was demonstrated in culture as evidenced by the ability of the steroid to prevent regression in explants derived from prostates of Copenhagen rats. The in vivo effects of Oe2 on the prostate weights could not be explained by differences in specific androgen or oestrogen receptor contents or in testosterone (T) metabolism. However, the prostates of younger Copenhagen rats differed from those of all other groups in three respects: 1) they contained high levels of oestramustine binding protein (OeBP), 2) they had the highest amount of uptake of radioactivity into the nuclear residue, and 3) their histological picture was characterized by diffuse stromal architecture having the appearance of oedematous tissue.

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THE EFFECT OF OESTRADIOL-3N-BIS-(2-CHLOROETHYL)CARBAMATE-17β-PHOSPHATE (ESTRACYT®) ON THE 5α-REDUCTASE IN THE RAT VENTRAL PROSTATE

Acta Endocrinologica ◽

10.1530/acta.0.0800188 ◽

1975 ◽

Vol 80 (1) ◽

pp. 188-198 ◽

Cited By ~ 7

Author(s):

Per Aage Høisaeter

Keyword(s):

Reductase Activity ◽

Inhibitory Effect ◽

Term Treatment ◽

Significant Inhibition ◽

Ventral Prostate ◽

Rat Ventral Prostate ◽

Long Term Treatment

ABSTRACT The ventral prostate of the rat both after in vitro incubation and in vivo experiments was found to contain appreciable 5α-reductase activity, whilst a very low activity was registered in the diaphragm and liver. Neither Estracyt® nor LEO275 (Estracyt® without the phosphate group in position 17 of the oestradiol moiety) had an inhibitory effect on the enzyme activity after in vitro incubation but equivalent amounts of oestradiol-17β and oestradiol-17β-phosphate significantly reduced 5α-reductase activity. When Estracyt® was injected in vivo no influence on activity was registered in "short term" experiments while a significant inhibition was found after "long term" treatment in vivo. Possible explanations for this "long term" effect of Estracyt® on 5α-reductase activity are discussed.

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Promising Anti-stroke Signature of Voglibose: Investigation through In- Silico Molecular Docking and Virtual Screening in In-Vivo Animal Studies

Current Gene Therapy ◽

10.2174/1566523220999200726225457 ◽

2020 ◽

Vol 20 (3) ◽

pp. 223-235

Author(s):

Pooja Shah ◽

Vishal Chavda ◽

Snehal Patel ◽

Shraddha Bhadada ◽

Ghulam Md. Ashraf

Keyword(s):

Molecular Docking ◽

Virtual Screening ◽

In Silico ◽

Reductase Activity ◽

Infarct Volume ◽

Docking Studies ◽

Serum Glucose ◽

In Vivo Studies ◽

In Silico Molecular Docking

Background: Postprandial hyperglycemia considered to be a major risk factor for cerebrovascular complications. Objective: The current study was designed to elucidate the beneficial role of voglibose via in-silico in vitro to in-vivo studies in improving the postprandial glycaemic state by protection against strokeprone type 2 diabetes. Material and Methods: In-Silico molecular docking and virtual screening were carried out with the help of iGEMDOCK+ Pymol+docking software and Protein Drug Bank database (PDB). Based on the results of docking studies, in-vivo investigation was carried out for possible neuroprotective action. T2DM was induced by a single injection of streptozotocin (90mg/kg, i.v.) to neonates. Six weeks after induction, voglibose was administered at the dose of 10mg/kg p.o. for two weeks. After eight weeks, diabetic rats were subjected to middle cerebral artery occlusion, and after 72 hours of surgery, neurological deficits were determined. The blood was collected for the determination of serum glucose, CK-MB, LDH and lipid levels. Brains were excised for determination of brain infarct volume, brain hemisphere weight difference, Na+-K+ ATPase activity, ROS parameters, NO levels, and aldose reductase activity. Results: In-silico docking studies showed good docking binding score for stroke associated proteins, which possibly hypotheses neuroprotective action of voglibose in stroke. In the present in-vivo study, pre-treatment with voglibose showed a significant decrease (p<0.05) in serum glucose and lipid levels. Voglibose has shown significant (p<0.05) reduction in neurological score, brain infarct volume, the difference in brain hemisphere weight. On biochemical evaluation, treatment with voglibose produced significant (p<0.05) decrease in CK-MB, LDH, and NO levels in blood and reduction in Na+-K+ ATPase, oxidative stress, and aldose reductase activity in brain homogenate. Conclusion: In-silico molecular docking and virtual screening studies and in-vivo studies in MCAo induced stroke, animal model outcomes support the strong anti-stroke signature for possible neuroprotective therapeutics.

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STEM-26. SMALL MOLECULE DRUG CBL0137 INHIBITS THE FACT COMPLEX AND SENSITIZES GLIOBLASTOMA CANCER STEM CELLS TO RADIOTHERAPY

Neuro-Oncology ◽

10.1093/neuonc/noaa215.843 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii201-ii202

Author(s):

Miranda Tallman ◽

Abigail Zalenski ◽

Amanda Deighen ◽

Morgan Schrock ◽

Sherry Mortach ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

High Rate ◽

Orthotopic Model ◽

In Vivo Models ◽

Specific Cytotoxicity ◽

Treatment Paradigm ◽

Cancer Agent

Abstract Glioblastoma (GBM) is a malignant brain tumor with nearly universal recurrence. GBM cancer stem cells (CSCs), a subpopulation of radio- and chemo-resistant cancer cells capable of self-renewal, contribute to the high rate of recurrence. The anti-cancer agent, CBL0137, inhibits the FACT (facilitates chromatin transcription) complex leading to cancer cell specific cytotoxicity. Here, we show that CBL0137 sensitized GBM CSCs to radiotherapy using both in vitro and in vivo models. Treatment of CBL0137 combined with radiotherapy led to increased DNA damage in GBM patient specimens and failure to resolve the damage led to decreased cell viability. Using clonogenic assays, we confirmed that CBL0137 radiosensitized the CSCs. To validate that combination therapy impacted CSCs, we used an in vivo subcutaneous model and showed a decrease in the frequency of cancer stem cells present in tumors as well as decreased tumor volume. Using an orthotopic model of GBM, we confirmed that treatment with CBL0137 followed by radiotherapy led to significantly increased survival compared to either treatment alone. Radiotherapy remains a critical component of patient care for GBM, even though there exists a resistant subpopulation. Radio-sensitizing agents, including CBL0137, pose an exciting treatment paradigm to increase the efficacy of irradiation, especially by inclusively targeting CSCs.

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Targeting the Extracellular Matrix in Abdominal Aortic Aneurysms Using Molecular Imaging Insights

International Journal of Molecular Sciences ◽

10.3390/ijms22052685 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2685

Author(s):

Lisa Adams ◽

Julia Brangsch ◽

Bernd Hamm ◽

Marcus R. Makowski ◽

Sarah Keller

Keyword(s):

Extracellular Matrix ◽

Molecular Imaging ◽

Molecular Targets ◽

Abdominal Aortic Aneurysms ◽

Aortic Aneurysms ◽

Type I ◽

Target Tissue ◽

Pharmacologic Treatment ◽

Abdominal Aortic

This review outlines recent preclinical and clinical advances in molecular imaging of abdominal aortic aneurysms (AAA) with a focus on molecular magnetic resonance imaging (MRI) of the extracellular matrix (ECM). In addition, developments in pharmacologic treatment of AAA targeting the ECM will be discussed and results from animal studies will be contrasted with clinical trials. Abdominal aortic aneurysm (AAA) is an often fatal disease without non-invasive pharmacologic treatment options. The ECM, with collagen type I and elastin as major components, is the key structural component of the aortic wall and is recognized as a target tissue for both initiation and the progression of AAA. Molecular imaging allows in vivo measurement and characterization of biological processes at the cellular and molecular level and sets forth to visualize molecular abnormalities at an early stage of disease, facilitating novel diagnostic and therapeutic pathways. By providing surrogate criteria for the in vivo evaluation of the effects of pharmacological therapies, molecular imaging techniques targeting the ECM can facilitate pharmacological drug development. In addition, molecular targets can also be used in theranostic approaches that have the potential for timely diagnosis and concurrent medical therapy. Recent successes in preclinical studies suggest future opportunities for clinical translation. However, further clinical studies are needed to validate the most promising molecular targets for human application.

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THE INFLUENCE OF SERUM ON THE UPTAKE, CONVERSION AND ACTION OF DIHYDROTESTOSTERONE IN RAT PROSTATE GLANDS IN ORGAN CULTURE

Journal of Endocrinology ◽

10.1677/joe.0.0640289 ◽

1975 ◽

Vol 64 (2) ◽

pp. 289-297 ◽

Cited By ~ 17

Author(s):

ILSE LASNITZKI ◽

HILARY R. FRANKLIN

Keyword(s):

Organ Culture ◽

Horse Serum ◽

Target Tissue ◽

Serum Free Medium ◽

Free Medium ◽

Serum Free ◽

Human Pregnancy ◽

Human Male ◽

Columnar Cells

SUMMARY The influence of serum on the uptake, conversion and action of dihydrotestosterone in relation to the sex steroid binding protein, TeBG, has been investigated in rat ventral prostates in organ culture. The organs were incubated with [1,2-3H]dihydrotestosterone in: (1) serum-free medium, (2) horse serum, foetal and newborn bovine serum or (3) human male and human pregnancy serum. With all sera the uptake of dihydrotestosterone fell with rising serum concentration, at first steeply and then more gradually. At the same concentration, the uptake was significantly lower in explants incubated with human pregnancy serum than in those kept with human male serum. The conversion of dihydrotestosterone to androstanediol followed the same pattern and less androstanediol was formed in the presence of pregnancy serum. Since pregnancy serum contains higher amounts of TeBG than male serum, the lowered uptake suggests that only the free hormone was available to the target organ. Addition of unlabelled dihydrotestosterone resulted in a higher uptake than that measured in explants incubated with the labelled steroid only. The effect of the human sera on uptake and conversion was correlated with the androgenic activity of dihydrotestosterone applied at physiological concentrations and expressed as the percentage of secretory columnar cells present. The degree of maintenance closely corresponded to the uptake of the hormone. In serum-free medium, the number of columnar cells approached the values found in vivo, with male serum their number, though reduced, was still substantial, with pregnancy serum it was extremely low. It is concluded that the amounts of TeBG present in serum regulate the supply of the hormone to the target tissue and thus control its biological action.

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A novel phosphoramide compound, DCZ0805, shows potent anti-myeloma activity via the NF-κB pathway

Cancer Cell International ◽

10.1186/s12935-021-01973-1 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Xuejie Gao ◽

Bo Li ◽

Anqi Ye ◽

Houcai Wang ◽

Yongsheng Xie ◽

...

Keyword(s):

Mouse Model ◽

Tumor Burden ◽

High Rate ◽

Cell Counting ◽

Tumor Xenograft ◽

Luciferase Reporter ◽

Therapeutic Effects ◽

Xenograft Mouse Model

Abstract Background Multiple myeloma (MM) is a highly aggressive and incurable clonal plasma cell disease with a high rate of recurrence. Thus, the development of new therapies is urgently needed. DCZ0805, a novel compound synthesized from osalmide and pterostilbene, has few observed side effects. In the current study, we intend to investigate the therapeutic effects of DCZ0805 in MM cells and elucidate the molecular mechanism underlying its anti-myeloma activity. Methods We used the Cell Counting Kit-8 assay, immunofluorescence staining, cell cycle assessment, apoptosis assay, western blot analysis, dual-luciferase reporter assay and a tumor xenograft mouse model to investigate the effect of DCZ0805 treatment both in vivo and in vitro. Results The results showed that DCZ0805 treatment arrested the cell at the G0/G1 phase and suppressed MM cells survival by inducing apoptosis via extrinsic and intrinsic pathways. DCZ0805 suppressed the NF-κB signaling pathway activation, which may have contributed to the inhibition of cell proliferation. DCZ0805 treatment remarkably reduced the tumor burden in the immunocompromised xenograft mouse model, with no obvious toxicity observed. Conclusion The findings of this study indicate that DCZ0805 can serve as a novel therapeutic agent for the treatment of MM.

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Fibrillar pharmacology of functionalized nanocellulose

Scientific Reports ◽

10.1038/s41598-020-79592-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sam Wong ◽

Simone Alidori ◽

Barbara P. Mello ◽

Bryan Aristega Almeida ◽

David Ulmert ◽

...

Keyword(s):

Aspect Ratio ◽

Fluorescent Dyes ◽

Pharmacokinetic Profile ◽

Water Soluble ◽

Size Exclusion ◽

Hydroxyl Groups ◽

Target Tissue ◽

Exclusion Chromatography ◽

Specific Accumulation

AbstractCellulose nanocrystals (CNC) are linear organic nanomaterials derived from an abundant naturally occurring biopolymer resource. Strategic modification of the primary and secondary hydroxyl groups on the CNC introduces amine and iodine group substitution, respectively. The amine groups (0.285 mmol of amine per gram of functionalized CNC (fCNC)) are further reacted with radiometal loaded-chelates or fluorescent dyes as tracers to evaluate the pharmacokinetic profile of the fCNC in vivo. In this way, these nanoscale macromolecules can be covalently functionalized and yield water-soluble and biocompatible fibrillar nanoplatforms for gene, drug and radionuclide delivery in vivo. Transmission electron microscopy of fCNC reveals a length of 162.4 ± 16.3 nm, diameter of 11.2 ± 1.52 nm and aspect ratio of 16.4 ± 1.94 per particle (mean ± SEM) and is confirmed using atomic force microscopy. Size exclusion chromatography of macromolecular fCNC describes a fibrillar molecular behavior as evidenced by retention times typical of late eluting small molecules and functionalized carbon nanotubes. In vivo, greater than 50% of intravenously injected radiolabeled fCNC is excreted in the urine within 1 h post administration and is consistent with the pharmacological profile observed for other rigid, high aspect ratio macromolecules. Tissue distribution of fCNC shows accumulation in kidneys, liver, and spleen (14.6 ± 6.0; 6.1 ± 2.6; and 7.7 ± 1.4% of the injected activity per gram of tissue, respectively) at 72 h post-administration. Confocal fluorescence microscopy reveals cell-specific accumulation in these target tissue sinks. In summary, our findings suggest that functionalized nanocellulose can be used as a potential drug delivery platform for the kidneys.

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Inhibition of Glycolysis Suppresses Cell Proliferation and Tumor Progression In Vivo: Perspectives for Chronotherapy

International Journal of Molecular Sciences ◽

10.3390/ijms22094390 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4390

Author(s):

Jana Horváthová ◽

Roman Moravčík ◽

Miroslava Matúšková ◽

Vladimír Šišovský ◽

Andrej Boháč ◽

...

Keyword(s):

Cell Proliferation ◽

Tumor Progression ◽

Umbilical Vein ◽

Cell Metabolism ◽

High Rate ◽

Ki 67 ◽

Immunodeficient Mice ◽

Inhibition Of Glycolysis

A high rate of glycolysis is considered a hallmark of tumor progression and is caused by overexpression of the enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3). Therefore, we analyzed the possibility of inhibiting tumor and endothelial cell metabolism through the inhibition of PFKFB3 by a small molecule, (E)-1-(pyridin-4-yl)-3-(quinolin-2-yl)prop-2-en-1-one (PFK15), as a promising therapy. The effects of PFK15 on cell proliferation and apoptosis were analyzed on human umbilical vein endothelial cells (HUVEC) and the human colorectal adenocarcinoma cell line DLD1 through cytotoxicity and proliferation assays, flow cytometry, and western blotting. The results showed that PFK15 inhibited the proliferation of both cell types and induced apoptosis with decreasing the Bcl-2/Bax ratio. On the basis of the results obtained from in vitro experiments, we performed a study on immunodeficient mice implanted with DLD1 cells. We found a reduced tumor mass after morning PFK15 treatment but not after evening treatment, suggesting circadian control of underlying processes. The reduction in tumor size was related to decreased expression of Ki-67, a marker of cell proliferation. We conclude that inhibition of glycolysis can represent a promising therapeutic strategy for cancer treatment and its efficiency is circadian dependent.

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Experimental Horizontal Transmission of Enterospora nucleophila (Microsporea: Enterocytozoonidae) in Gilthead Sea Bream (Sparus aurata)

Animals ◽

10.3390/ani11020362 ◽

2021 ◽

Vol 11 (2) ◽

pp. 362

Author(s):

Amparo Picard-Sánchez ◽

M. Carla Piazzon ◽

Itziar Estensoro ◽

Raquel Del Pozo ◽

Nahla Hossameldin Ahmed ◽

...

Keyword(s):

Sparus Aurata ◽

Horizontal Transmission ◽

Infected Fish ◽

Gilthead Sea Bream ◽

Sea Bream ◽

Target Tissue ◽

Farmed Fish ◽

Infection Level ◽

Available Information

Enterospora nucleophila is a microsporidian enteroparasite that infects mainly the intestine of gilthead sea bream (Sparus aurata), leading to an emaciative syndrome. Thus far, the only available information about this infection comes from natural outbreaks in farmed fish. The aim of the present study was to determine whether E. nucleophila could be transmitted horizontally using naturally infected fish as donors, and to establish an experimental in vivo procedure to study this host–parasite model without depending on natural infections. Naïve fish were exposed to the infection by cohabitation, effluent, or intubated either orally or anally with intestinal scrapings of donor fish in four different trials. We succeeded in detecting parasite in naïve fish in all the challenges, but the infection level and the disease signs were always milder than in donor fish. The parasite was found in peripheral blood of naïve fish at 4 weeks post-challenge (wpc) in oral and effluent routes, and up to 12 wpc in the anal transmission trial. Molecular diagnosis detected E. nucleophila in other organs besides intestine, such as gills, liver, stomach or heart, although the intensity was not as high as in the target tissue. The infection tended to disappear through time in all the challenge routes assayed, except in the anal infection route.

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