scholarly journals Ellagitannins – Compounds From Pomegranate as Possible Effector in Steroidogenesis of Rabbit Ovaries

2015 ◽  
pp. 583-585 ◽  
Author(s):  
D. PACKOVA ◽  
A. A. CARBONELL-BARRACHINA ◽  
A. KOLESAROVA
Keyword(s):  
Steroid Hormones ◽  
Mature Female ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Progesterone Secretion ◽  
New Zealand White Rabbits ◽  
17Β Estradiol ◽  
Dose Dependent ◽  
Sexually Mature

This study has observed possible effect of ellagitannins – compounds from pomegranate on process of steroidogenesis in ovaries. The aim of the study was to investigate the possible effect of punicalagin on secretion of steroid hormones – progesterone, androstenedione, testosterone and 17β-estradiol by ovarian fragments of rabbits in vitro. Ovarian fragments from sexually mature female New Zealand white rabbits (n=20) were incubated without (control group) or with punicalagin at various doses 1, 10 and 100 μg.ml−1 for 24 h. Hormones were evaluated by ELISA (The Enzyme-Linked Immunosorbent Assay). Data showed that progesterone and 17β-estradiol (but not androstenedione and testosterone) release by rabbit ovarian fragments was significantly affected by punicalagin addition at various doses. Punicalagin (at 100 μg.ml−1) significantly (P<0.05) increased progesterone secretion. On the other hand, the release of 17β-estradiol was significantly (P<0.005) decreased by punicalagin addition (at 10 μg.ml−1). Our results suggest that punicalagin could have dose-dependent impact on secretion of steroid hormones progesterone and 17β-estradiol by rabbit ovarian fragments and it may be effector in process of ovarian steroidogenesis.

scholarly journals Steroid hormones regulate sperm–oviduct interactions in the bovine

Reproduction ◽  
2017 ◽  
Vol 154 (4) ◽  
pp. 497-508 ◽  
Author(s):  
Julie Lamy ◽  
Emilie Corbin ◽  
Marie-Claire Blache ◽  
Anastasiia S Garanina ◽  
Rustem Uzbekov ◽  
...  
Keyword(s):  
Steroid Hormones ◽  
Hormonal Regulation ◽  
Control Group ◽  
Dependent Manner ◽  
Dynamic Changes ◽  
17Β Estradiol ◽  
Sperm Reservoir ◽  
Releasing Effect ◽  
Dose Dependent

After insemination in the cow, a sperm reservoir is formed within the oviducts, allowing the storage and then progressive release of spermatozoa toward the ovulated oocyte. In order to investigate the hormonal regulation of these events in vitro, the ovarian steroids 17β-estradiol (E2) and progesterone (P4) were added at various concentrations to monolayers of bovine oviduct epithelial cells (BOEC) before or during co-incubation with spermatozoa. Main findings demonstrate that (1) a 18-h pretreatment of BOEC with 100 pg/mL and 100 ng/mL of E2 decreased by 25% the ability of BOEC to bind spermatozoa after 10 min, and for the highest dose of E2, 60 min of co-incubation; (2) P4 at concentrations of 10, 100 and 1000 ng/mL induced the release within 60 min of 32–47% of bound spermatozoa from BOEC; this sperm-releasing effect was maintained after a 18-h pretreatment of BOEC with 100 pg/mL of E2; (3) E2 in concentrations above 100 pg/mL inhibited the releasing effect of P4 on bound sperm in a dose-dependent manner; (4) spermatozoa bound to BOEC, then released from BOEC by the action of P4-induced higher cleavage and blastocyst rates after in vitro fertilization than the control group. These results support the hypothesis that the dynamic changes in steroid hormones around the time of ovulation regulate the formation of the sperm reservoir and the timed delivery of capacitated spermatozoa to the site of fertilization.

scholarly journals In Vitro Assessment of the Impact of Nickel on the Viability and Steroidogenesis in the Human Adrenocortical Carcinoma (NCI-H295R) Cell Line

2020 ◽  
pp. 871-883
Author(s):  
Norbert LUKAC ◽  
Z FORGACS ◽  
H DURANOVA ◽  
T JAMBOR ◽  
J ZEMANOVA ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Adrenocortical Carcinoma ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Cytotoxic Action ◽  
Disruptive Effect ◽  
17Β Estradiol ◽  
The Impact

Nickel is a ubiquitous environmental pollutant, which has various effects on reproductive endocrinology. In this study, human adrenocortical carcinoma (NCI-H295R) cell line was used as an in vitro biological model to study the effect of nickel chloride (NiCl2) on the viability and steroidogenesis. The cells were exposed to different concentrations (3.90; 7.80; 15.60; 31.20; 62.50; 125; 250 and 500 μM) of NiCl2 and compared with control group (culture medium without NiCl2). The cell viability was measured by the metabolic activity assay. Production of sexual steroid hormones was quantified by enzyme linked immunosorbent assay. Following 48 h culture of the cells in the presence of NiCl2 a dose-dependent depletion of progesterone release was observed even at the lower concentrations. In fact, lower levels of progesterone were detected in groups with higher doses (≥125 μM) of NiCl2 (P<0.01), which also elicited cytotoxic action. A more prominent decrease in testosterone production (P<0.01) was also noted in comparison to that of progesterone. On the other hand, the release of 17β-estradiol was substantially increased at low concentrations (3.90 to 62.50 μM) of NiCl2. The cell viability remained relatively unaltered up to 125 μM (P>0.05) and slightly decreased from 250 μM of NiCl2 (P<0.05). Our results indicate endocrine disruptive effect of NiCl2 on the release of progesterone and testosterone in the NCI-H295R cell line. Although no detrimental effect of NiCl2 (≤62.50 μM) could be found on 17β-estradiol production, its toxicity may reflect at other points of the steroidogenic pathway.

scholarly journals Effect of Synchronous Versus Sequential Regimens on the Pharmacokinetics and Biodistribution of Regorafenib with Irradiation

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 386
Author(s):  
Tung-Hu Tsai ◽  
Yu-Jen Chen ◽  
Li-Ying Wang ◽  
Chen-Hsi Hsieh
Keyword(s):  
Stereotactic Body Radiation Therapy ◽  
In Vivo Studies ◽  
Control Group ◽  
High Dose ◽  
Dependent Manner ◽  
Hep G2 ◽  
Time Curve ◽  
Dose Dependent

This study was performed to evaluate the interaction between conventional or high-dose radiotherapy (RT) and the pharmacokinetics (PK) of regorafenib in concurrent or sequential regimens for the treatment of hepatocellular carcinoma. Concurrent and sequential in vitro and in vivo studies of irradiation and regorafenib were designed. The interactions of RT and regorafenib in vitro were examined in the human hepatoma Huh-7, HA22T and Hep G2 cell lines. The RT–PK phenomenon and biodistribution of regorafenib under RT were confirmed in a free-moving rat model. Regorafenib inhibited the viability of Huh-7 cells in a dose-dependent manner. Apoptosis in Huh-7 cells was enhanced by RT followed by regorafenib treatment. In the concurrent regimen, RT decreased the area under the concentration versus time curve (AUC)regorafenib by 74% (p = 0.001) in the RT2 Gy × 3 fraction (f’x) group and by 69% (p = 0.001) in the RT9 Gy × 3 f’x group. The AUCregorafenib was increased by 182.8% (p = 0.011) in the sequential RT2Gy × 1 f’x group and by 213.2% (p = 0.016) in the sequential RT9Gy × 1 f’x group. Both concurrent regimens, RT2Gy × 3 f’x and RT9Gy × 3 f’x, clearly decreased the biodistribution of regorafenib in the heart, liver, lung, spleen and kidneys, compared to the control (regorafenib × 3 d) group. The concurrent regimens, both RT2Gy × 3 f’x and RT9Gy × 3 f’x, significantly decreased the biodistribution of regorafenib, compared with the control group. The PK of regorafenib can be modulated both by off-target irradiation and stereotactic body radiation therapy (SBRT).

scholarly journals The Effect of HT-2 Toxin on Ovarian Steroidogenesis and Its Response to IGF-I, Leptin and Ghrelin in Rabbits

2017 ◽  
pp. 705-708 ◽  
Author(s):  
A. KOLESAROVA ◽  
N. MARUNIAKOVA ◽  
A. KADASI ◽  
M. HALENAR ◽  
M. MARAK ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fusarium Species ◽  
Endocrine System ◽  
Control Group ◽  
Trichothecene Mycotoxin ◽  
Ovarian Steroidogenesis ◽  
Estradiol Secretion ◽  
Igf I ◽  
17Β Estradiol

T-2 toxin and its metabolite HT-2 toxin are one of the most toxic mycotoxins of type A-trichothecenes, which are produced mainly by Fusarium species. Therefore, study of Fusarium toxins T-2 toxin and HT-2 toxin is an essential issue because they could also play role in failures of reproductive functions as well as endocrine system of domestic animals. Assessment of the effect of A-trichothecene mycotoxin HT-2 toxin alone or combined with insulin-like growth factor (IGF-I), leptin and ghrelin on estradiol secretion by rabbit ovarian fragments in vitro was done. Rabbit ovarian fragments were incubated without (control group) or with HT-2 toxin, or its combinations with IGF-I, leptin and ghrelin at various concentrations for 24 h. Secretion of 17β-estradiol was determined by ELISA. Firstly, HT-2 toxin at the doses 10 and 100 ng.ml-1, but not at 1 ng.ml-1 decreased 17β-estradiol secretion by ovarian fragments. Secondly, 17β-estradiol secretion was not affected by HT-2 toxin exposure combined with growth factor IGF-I, metabolic hormones leptin and ghrelin. In conclusion, HT-2 toxin has potent direct dose-dependent effects on ovarian steroidogenesis in rabbits. These direct effects of HT-2 mycotoxin on ovarian steroidogenesis could impact negatively on the reproductive performance of rabbits.

Production and utilization of monoclonal antibodies to human/rat corticotrophin-releasing factor-41

1990 ◽  
Vol 5 (2) ◽  
pp. 159-166 ◽  
Author(s):  
N. G. N. Milton ◽  
E. W. Hillhouse ◽  
S. A. Nicholson ◽  
C. H. Self ◽  
A. M. McGregor
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Corticotrophin Releasing Factor ◽  
Tissue Extracts ◽  
Rat Pituitary ◽  
Hypothalamic Tissue ◽  
Dose Dependent

ABSTRACT Murine monoclonal antibodies against human/rat corticotrophin-releasing factor-41 (CRF-41) were produced and characterized for use in the immunological and biological characterization of CRF-41. Spleen cells from BALB/c mice immunized with CRF-41 conjugated to bovine γ-globulin were fused with a BALB/c-derived non-secretor X-63 myeloma line. Hybridomas were selected for CRF antibody production by enzyme-linked immunosorbent assay, and positive hybridomas cloned twice. Three monoclonal antibodies were obtained (KCHMB001, KCHMB002 and KCHMB003) and characterized as IgG1, IgG1 and IgG2a isotypes respectively, with affinity constants for rat CRF-41 of 30, 53 and 34 nmol/l respectively. All three monoclonal antibodies recognize an epitope contained between residues 34 and 41 of the human/rat sequence. The antibodies were able to neutralize the ACTH-releasing activity of rat CRF-41, applied to rat pituitary fragments in vitro, in a dose-dependent manner. Isoelectric focusing showed that KCHMB 003 detected bands of synthetic rat CRF-41 and rat [Met(O)21,38]-CRF-41 at pH 7·1 and 6·8 respectively. Use of KCHMB003 in a two-site enzyme-amplified immunoassay showed that this antibody recognizes both synthetic rat CRF-41 and immunoreactive CRF-41 in rat hypothalamic tissue extracts.

scholarly journals THE TOXICITY OF MAULI BANANA (MUSA ACUMINATA) STEM WATER EXTRACT ON BONE MARROW MESENCHYMAL STEM CELL IN VITRO

2019 ◽  
pp. 184-186
Author(s):  
AMY NINDIA ◽  
DIDIT ASPRIYANTO ◽  
MAHARANI LAILLYZA APRIASARI ◽  
SELVIANA RIZKY
Keyword(s):  
Growth Factor ◽  
Water Extract ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Cell Culture Process ◽  
Control Group Design ◽  
Water Solvent ◽  
Stem Water ◽  
Banana Stem

Objective: Since mesenchymal stem cells (MSC) can differentiate into bone, cementum, and periodontal ligament, they can be used to treat aggressiveperiodontitis. The limited number of MSCs requires replenishment of growth factor in the cell culture process. Since growth factor is quite expensive,an alternative material is needed. Mauli banana stem has antioxidant and immunomodulatory properties. Methanol extract of Mauli banana stem isknown to be toxic toward MSCs; therefore, another solvent with a non-toxic effect is needed, such as a water solvent. We analyzed the toxicity of Maulibanana stem water extract on MSC in vitro.Methods: In this laboratory experimental (true experimental) study with a Post-test Only Control Group Design, MSC cultures were treated withMauli banana stem water extract at 10, 20, 40, 60, 80, and 100 mg/mL dosages. One group without any treatment served as a control group and onewas a media control group. Each group was incubated for 24 h and then was given 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidereagent and analyzed by an enzyme-linked immunosorbent assay (ELISA) reader.Results: One-way analysis of variance showed a significant difference.Conclusion: Mauli banana stem water extracts at 10, 20, 40, and 60 mg/mL were not toxic toward MSC in vitro, while dosages of 80 and 100 mg/mLdosage were toxic.

scholarly journals Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins

2017 ◽  
Vol 2017 ◽  
pp. 1-14
Author(s):  
Mayra Rodríguez-Rodríguez ◽  
Rafael Herrera-Esparza ◽  
Juan-José Bollain y Goytia ◽  
María-Elena Pérez-Pérez ◽  
Deyanira Pacheco-Tovar ◽  
...  
Keyword(s):  
Salivary Glands ◽  
Posttranslational Modifications ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Peptidylarginine Deiminase ◽  
Reporter Protein ◽  
Ductal Cells ◽  
Arginine Residues

The goal of the present study was to determine whether peptidylarginine deiminase PAD2 and PAD4 enzymes are present in Balb/c mouse salivary glands and whether they are able to citrullinate Ro and La ribonucleoproteins. Salivary glands from Balb/c mice were cultured in DMEM and supplemented with one of the following stimulants: ATP, LPS, TNF, IFNγ, or IL-6. A control group without stimulant was also evaluated. PAD2, PAD4, citrullinated peptides, Ro60, and La were detected by immunohistochemistry and double immunofluorescence. PAD2 and PAD4 mRNAs and protein expression were detected by qPCR and Western blot analysis. PAD activity was assessed using an antigen capture enzyme-linked immunosorbent assay. LPS, ATP, and TNF triggered PAD2 and PAD4 expression; in contrast, no expression was detected in the control group (p<0.001). PAD transcription slightly increased in response to stimulation. Additionally, PAD2/4 activity modified the arginine residues of a reporter protein (fibrinogen) in vitro. PADs citrullinated Ro60 and La ribonucleoproteins in vivo. Molecular stimulants induced apoptosis in ductal cells and the externalization of Ro60 and La ribonucleoproteins onto apoptotic membranes. PAD enzymes citrullinate Ro and La ribonucleoproteins, and this experimental approach may facilitate our understanding of the role of posttranslational modifications in the pathophysiology of Sjögren’s syndrome.

Protein-free phospholipid emulsion treatment improved cardiopulmonary function and survival in porcine sepsis

2003 ◽  
Vol 284 (2) ◽  
pp. R550-R557 ◽  
Author(s):  
Roy D. Goldfarb ◽  
Thomas S. Parker ◽  
Daniel M. Levine ◽  
Dana Glock ◽  
Imran Akhter ◽  
...  
Keyword(s):  
Density Lipoprotein ◽  
Fibrin Clot ◽  
Control Group ◽  
High Dose ◽  
Dependent Manner ◽  
Serum Lipoproteins ◽  
Treatment Groups ◽  
Tnf Α ◽  
Dose Dependent

Lipoprotein phospholipid (PL) plays a major role in neutralization of endotoxin. This study tested the hypothesis that prophylactic administration of a PL-enriched emulsion (PRE), which augments PL content of serum lipoproteins and neutralizes endotoxin in vitro, would preserve cardiovascular function and improve survival in porcine septic peritonitis. A control group was compared with low-, mid-, and high-dose treatment groups that received PRE by primed continuous infusion for 48 h. A fibrin clot containing live Escherichia coli 0111.B4 was implanted intraperitoneally 30 min after the priming dose. Survival increased in a dose-dependent manner and was correlated with serum PL. Infused PL was associated with high-density lipoprotein in the low-dose group and all serum lipoproteins at higher doses. Treatment significantly lowered serum endotoxin and tumor necrosis factor (TNF)-α, preserved cardiac output and ejection fraction, and attenuated increases in systemic and pulmonary vascular resistances. This study demonstrated that augmentation of lipoprotein PL via administration of PRE improved survival and offered a novel therapeutic approach to sepsis.

scholarly journals Xylazine regulates the release of glycine and aspartic acid in rat brain

2018 ◽  
Vol 62 (1) ◽  
pp. 121-128
Author(s):  
Yi-Ming Zhang ◽  
Dong-Xu Yu ◽  
Bai-Shuang Yin ◽  
Xin-Ran Li ◽  
Li-Na Li ◽  
...  
Keyword(s):  
Aspartic Acid ◽  
Brain Area ◽  
Brain Regions ◽  
Inhibitory Effect ◽  
Test Methods ◽  
Control Group ◽  
Clinical Anaesthesia ◽  
Induced Changes ◽  
Dose Dependent

AbstractIntroductionXylazine, a type of α2-adrenoceptors, is a commonly used drug in veterinary medicine. Xylazine-induced changes in the content of amino acid neurotransmitters – glycine (Gly) and aspartic acid (Asp), in different brain regions and neurons were studied.Material and MethodsWistar rats were administered 50 mg/kg or 70 mg/kg of xylazine by intraperitoneal injection. In addition, in vitro experiments were conducted, in which neurons were treated with 15 μg/mL, 25 μg/mL, 35μg/mL, and 45 μg/mL of xylazine. Test methods were based on the enzyme-linked immunosorbent assays (ELISA).ResultsDuring anaesthesia, Asp levels in each brain area were significantly lower compared to the control group. Except for the cerebrum, levels of Gly in other brain areas were significantly increased during the anaesthesia period. In vitro, xylazine-related neuron secretion of Gly increased significantly compared to the control group at 60 min and 90 min. Moreover, xylazine caused a significant decrease in the levels of Asp secreted by neurons at 20 min, but gradually returned to the level of the control group.ConclusionThe data showed that during anaesthesia the overall levels of Asp decreased and overall levels of Gly increased. In addition, the inhibitory effect of xylazine on Asp and the promotion of Gly were dose-dependent. Our data showed that different effects of xylazine on excitatory and inhibitory neurotransmitters provided a theoretical basis for the mechanism of xylazine activity in clinical anaesthesia.

scholarly journals In vitro inhibition of porcine cytochrome P450 by 17β-estradiol and 17α-estradiol

2011 ◽  
Vol 4 (2) ◽  
pp. 78-84 ◽  
Author(s):  
Galia Zamaratskaia ◽  
Martin Rasmussen ◽  
Isabelle Herbin ◽  
Bo Ekstrand ◽  
Vladimir Zlabek
Keyword(s):  
Cytochrome P450 ◽  
Inhibitory Effect ◽  
17Β Estradiol ◽  
Cyp2e1 Activity ◽  
High Concentrations ◽  
Testicular Steroids ◽  
Sexually Mature

In vitro inhibition of porcine cytochrome P450 by 17β-estradiol and 17α-estradiol Sexually mature pigs are known to possess high concentrations of testicular steroids, which have been shown to change the activities of cytochrome P450 in vitro. The aim of the present study was to evaluate the regulation of CYP1A and CYP2E1 activity by the steroids dihydrotestosterone (DHT), 3β-androstenol, 17β-estradiol and 17α-estradiol. Catalytic activities of 7-ethoxyresorufin O-deethylase (EROD) and 7-methoxyresorufin O-demethylase (MROD) were used as markers of CYP1A activities, while p-nitrophenol hydroxylase (PNPH) was used as a marker of CYP2E1 activities. Of the steroids tested, only 17β-estradiol and 17α-estradiol inhibited EROD and MROD activities. This inhibition was observed when a steroid concentration of 100 μM was used, while lower concentrations showed no inhibitory effect. PNPH activities were inhibited only by 100 μM of 17β-estradiol. The significance of these results in vivo is unknown because inhibition was only found when concentrations of estrogens higher than physiological levels were used. Nevertheless, the results provided further evidence on the important role of estrogens in regulation of porcine cytochrome P450 activities.

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