scholarly journals Bisphenols effects on Granulosa Cell-Cell Communication

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A484-A484
Author(s):  
Reem Sabry ◽  
Charlotte Apps ◽  
Cassidy Van Den Diepstraten ◽  
Laura A Favetta
Keyword(s):  
Cell Viability ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Preliminary Data ◽  
Low Dose ◽  
Cell Communication ◽  
Endocrine Disrupting Compounds ◽  
High Dose ◽  
Medium Dose ◽  
Cell Cell

Abstract Bisphenols are Endocrine Disrupting Compounds (EDCs) linked to negative fertility outcomes in humans. The most common one, bisphenol A (BPA) is a plasticizer that acts as an estrogen agonist. Alternatives such as BPS and BPF are becoming widespread and possess similar activities. The oocyte and nearby granulosa cells are particularly vulnerable to EDCs as they are dependent on hormone signaling. Granulosa-Granulosa cell communications are pivotal for proper development and are mediated by gap junction molecules, called connexins (Cxs). Cx37, Cx26, and Cx43 are crucial in follicular communication and are the focus of this study. Bisphenols can alter these genes which directly affects cell-cell communication and in turn proper development. This study uses an in vitro cell culture model of bovine granulosa cells as the bovine species is an excellent translational model for human reproductive toxicology. To determine optimal doses and exposure times, we performed a cytotoxic assay (CCK8) and treated cells with a low dose (0.5 μg/mL), a medium dose (5 μg/mL), the current reported LOAEL (50 μg/mL), and a high dose (0.5 mg/mL) of bisphenols for 1, 6, 12, 24, 48, and 72 hrs. The high dose was lethal at all time points for BPA/BPF, while BPS was toxic from 1 hrs (P=0.0016; n=4). The LOAEL dose was toxic for BPA at 6, 12, 24 hrs (P=0.018, 0.0021, 0.0042 respectively; n=7) and was lethal at 48/72 hrs. BPF at the LOAEL dose showed an increase in cell viability at 6 hrs which was significant at 12hrs (P=0.0119; n=5). This trend declined at 48/72 hrs but was not significant. BPS did not affect cell viability at any other dose and the medium dose did not affect viability for all bisphenols; however, the low dose of BPA/BPF showed a trend towards decreased viability at 48 hrs of treatment. Therefore, we looked at the effects of BPA/S/F at the low and LOAEL doses for 12/48 hrs. We used qPCR and western blotting to quantify transcripts and proteins of the three connexins. Our data show that Cx43 mRNA was significantly increased at the LOAEL dose for BPA/BPF at 12 hrs (p=0.02; n=6). Preliminary data on proteins indicate a decrease and increase in expression at 12hrs for BPA/BPF treatment, respectively, at the LOAEL dose. BPS shows no effect on Cx43. BPS low dose and BPA/BPF LOAEL doses at 12 and 48 hrs increase Cx37 mRNA. Preliminary data on the protein levels show a significant decrease of Cx37 protein at BPA low dose, but no changes after exposure to the other bisphenols. Lastly, Cx26 mRNA is significantly increased after BPA treatment at the LOAEL dose at 12 hrs (P<0.0001; n=5). This novel study investigates the effects of low/high doses and short-term/chronic exposure of bisphenols on cell-cell communication in granulosa cells and provides the basis for future studies on bisphenol toxicity in early development.

scholarly journals Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sai Kumar ◽  
Meeti Punetha ◽  
Bosco Jose ◽  
Jaya Bharati ◽  
Shivani Khanna ◽  
...  
Keyword(s):  
Cell Viability ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cell Function ◽  
Biological Effects ◽  
Capra Hircus ◽  
Gene Modulation ◽  
High Fecundity ◽  
Smad Signaling ◽  
Sheep Breeds

AbstractBMPs are multifunctional growth factors implicated in regulating the ovarian function as key intra-ovarian factors. Biological effects of BMPs are mediated through binding with membrane bound receptors like BMPR-IB and initiating downstream Smad signaling pathway. FecB mutation, regarded as a loss of function mutation in the BMPR-IB gene was identified in certain sheep breeds having high fecundity. Similar type of fecundity genes in goats have not been discovered so far. Hence, the current study was designed to investigate the effects of BMPR-IB gene modulation on granulosa cell function in goats. The BMPR-IB gene was knocked out using CRISPR-Cas technology in granulosa cells and cultured in vitro with BMP-4 stimulation for three different durations In addition, the FecB mutation was introduced in the BMPR-IB gene applying Easi-CRISPR followed by BMP-4/7 stimulation for 72 h. Steroidogenesis and cell viability were studied to explore the granulosa cell function on BMPR-IB gene modulation. BMPRs were found to be expressed stage specifically in granulosa cells of goats. Higher transcriptional abundance of R-Smads, LHR and FSHR indicating sensitisation of Smad signaling and increased gonadotropin sensitivity along with a significant reduction in the cell proliferation and viability was observed in granulosa cells upon BMPR-IB modulation. The inhibitory action of BMP-4/7 on P4 secretion was abolished in both KO and KI cells. Altogether, the study has revealed an altered Smad signaling, steroidogenesis and cell viability upon modulation of BMPR-IB gene in granulosa cells similar to that are documented in sheep breeds carrying the FecB mutation.

scholarly journals M211. NEUROPROTECTIVE EFFECT OF SHI-ZHEN-AN-SHEN-TANG, A CHINESE HERB FORMULA ON MICE EXPOSED TO CUPRIZONE

2020 ◽  
Vol 46 (Supplement_1) ◽  
pp. S216-S217
Author(s):  
Chao Ma ◽  
Yan Wu ◽  
Pei Chen ◽  
Yuan Jia ◽  
Dongqing Yin ◽  
...  
Keyword(s):  
Low Dose ◽  
Neuroprotective Effect ◽  
Chinese Herb ◽  
Large Body ◽  
High Dose ◽  
Neuregulin 1 ◽  
The Brain ◽  
Different Doses ◽  
Medium Dose

Abstract Background Our previous study indicated a therapeutic effect of Shi-Zhen-An-Shen-Tang (SZAST), a Chinese herb formula, on schizophrenia, but the related mechanism is unknown(citation). A large body of evidence suggests the important role of white matter of the brain in the pathophysiology of schizophrenia. This study was designed to evaluate the effect of SZAST on schizophrenia with demyelinated mice. Methods Male C57BL/6 mice were given mixed cuprizone (CPZ, a copper chelator, 0.2 %, w/w) rodent chow for six successive weeks to induce demyelination. During the last two weeks, mice were given an oral gavage of saline, or SZAST of three different doses (a low dose of 5.5g·kg-1·d-1, a medium dose of 8.24g·kg-1·d-1, or a high dose of 10.98 g·kg-1·d-1), or quetiapine, respectively. Behavioral tests were conducted after the last treatment. Meanwhile, the expression of myelin basic protein (MBP) and neuregulin-1(NRG1) in the brain was tested by immunohistochemistry staining or Western Blot. Results Mice exposed to CPZ for six weeks showed obvious schizophrenia-like behaviors, including lower nest-building activity, sensory gating activity, and higher locomotor activity. CPZ-fed mice also displayed a lower myelin density in the corpus callosum, hippocampus, and cerebral cortex and a reduction of MBP and NRG1 protein in the hippocampus compared with controls. Both quetiapine and SZAST significantly alleviated the abnormal schizophrenia-like behaviors and the impairment of myelin sheath in CPZ-fed mice, however, SZAST with medium dose showed better neuroprotective effect than the low dose or the high dose of SZAST. Furthermore, the expression of NRG1protein in the hippocampus was slightly, but not significantly increased in all SZAST-treated and quetiapine-treated groups. Discussion These results indicate that the neuroprotective effect of SZAST in demyelinated mice might partially relate to remyelination in the hippocampus in CPZ-fed mice.

scholarly journals Dosing of thromboprophylaxis and mortality in critically ill COVID-19 patients

Critical Care ◽  
2020 ◽  
Vol 24 (1) ◽  
Author(s):  
Sandra Jonmarker ◽  
Jacob Hollenberg ◽  
Martin Dahlberg ◽  
Otto Stackelberg ◽  
Jacob Litorell ◽  
...  
Keyword(s):  
Body Weight ◽  
Confidence Intervals ◽  
Critically Ill ◽  
Low Dose ◽  
Cox Proportional Hazards ◽  
High Dose ◽  
Thromboembolic Events ◽  
Risk Of Death ◽  
Dosing Strategy ◽  
Medium Dose

Abstract Background A substantial proportion of critically ill COVID-19 patients develop thromboembolic complications, but it is unclear whether higher doses of thromboprophylaxis are associated with lower mortality rates. The purpose of the study was to evaluate the association between initial dosing strategy of thromboprophylaxis in critically ill COVID-19 patients and the risk of death, thromboembolism, and bleeding. Method In this retrospective study, all critically ill COVID-19 patients admitted to two intensive care units in March and April 2020 were eligible. Patients were categorized into three groups according to initial daily dose of thromboprophylaxis: low (2500–4500 IU tinzaparin or 2500–5000 IU dalteparin), medium (> 4500 IU but < 175 IU/kilogram, kg, of body weight tinzaparin or > 5000 IU but < 200 IU/kg of body weight dalteparin), and high dose (≥ 175 IU/kg of body weight tinzaparin or ≥ 200 IU/kg of body weight dalteparin). Thromboprophylaxis dosage was based on local standardized recommendations, not on degree of critical illness or risk of thrombosis. Cox proportional hazards regression was used to estimate hazard ratios with corresponding 95% confidence intervals of death within 28 days from ICU admission. Multivariable models were adjusted for sex, age, body mass index, Simplified Acute Physiology Score III, invasive respiratory support, and initial dosing strategy of thromboprophylaxis. Results A total of 152 patients were included: 67 received low-, 48 medium-, and 37 high-dose thromboprophylaxis. Baseline characteristics did not differ between groups. For patients who received high-dose prophylaxis, mortality was lower (13.5%) compared to those who received medium dose (25.0%) or low dose (38.8%), p = 0.02. The hazard ratio of death was 0.33 (95% confidence intervals 0.13–0.87) among those who received high dose, and 0.88 (95% confidence intervals 0.43–1.83) among those who received medium dose, as compared to those who received low-dose thromboprophylaxis. There were fewer thromboembolic events in the high (2.7%) vs medium (18.8%) and low-dose thromboprophylaxis (17.9%) groups, p = 0.04. Conclusions Among critically ill COVID-19 patients with respiratory failure, high-dose thromboprophylaxis was associated with a lower risk of death and a lower cumulative incidence of thromboembolic events compared with lower doses. Trial registration Clinicaltrials.gov NCT04412304 June 2, 2020, retrospectively registered.

A Double-Blind, Randomized Study of Extended-Release Molindone for Impulsive Aggression in ADHD

2020 ◽  
pp. 108705472090908 ◽  
Author(s):  
Gianpiera Ceresoli-Borroni ◽  
Azmi Nasser ◽  
Toyin Adewole ◽  
Tesfaye Liranso ◽  
Jiahong Xu ◽  
...  
Keyword(s):  
Extended Release ◽  
Low Dose ◽  
Behavioral Therapy ◽  
Randomized Study ◽  
High Dose ◽  
Double Blind ◽  
Impulsive Aggression ◽  
Pediatric Populations ◽  
Dose Ranging ◽  
Medium Dose

Objective: To evaluate efficacy and safety of SPN-810 (extended-release molindone) in a Phase-2b, randomized, double-blind, placebo-controlled, dose-ranging study of children (6–12 years) with ADHD and persistent impulsive aggression (IA). Method: After lead-in, children were randomized to (a) placebo ( N = 31); (b) low-dose ( N = 29, 12/18 mg/day); (c) medium-dose ( N = 30, 24/36 mg/day); and (4) high-dose ( N = 31, 36/54 mg/day) groups. Treatment included ~2.5-week titration, 3-week maintenance, and 1-week tapering/conversion, alongside existing monotherapy (stimulants/nonstimulants) and behavioral therapy. The primary endpoint was change in Retrospective-Modified Overt Aggression Scale (R-MOAS) score at end of study, with safety monitored. Results: A total of 95 (78.5%) children completed the study. Aggression (R-MOAS) improved with low and medium doses (low dose: p = .031; medium dose: p = .024; high dose: p = .740). The most common adverse events were headache (10.0%), sedation (8.9%), and increased appetite (7.8%). Conclusion: These results suggest SPN-810 may be effective in reducing residual IA behaviors in children with ADHD. Research is still needed to support the benefit–risk profile of SPN-810 in pediatric populations.

Simulation of Transient Enhanced Diffusion in Silicon Taking into Account Ostwald Ripening of Defects

2002 ◽  
Vol 717 ◽  
Author(s):  
Masashi Uematsu
Keyword(s):  
Time Dependence ◽  
Diffusion Model ◽  
Low Dose ◽  
Ostwald Ripening ◽  
High Dose ◽  
Enhanced Diffusion ◽  
Annealing Conditions ◽  
Transient Enhanced Diffusion ◽  
High Dose Implantation ◽  
Medium Dose

AbstractThe transient enhanced diffusion (TED) of high-dose implanted P is simulated taking into account Ostwald ripening of end-of-range (EOR) defects. First, we integrated a basic diffusion model based on the simulation of in-diffusion, where no implanted damages are involved. Second, from low-dose implantation, we developed a model for TED due to {311} self-interstitial (I) clusters involving Ostwald ripening and the dissolution of {311} clusters. Third, from medium-dose implantation, we showed that P-I clusters should be taken into account, and during the diffusion, the clusters are dissolved to emit self-interstitials that also contribute to TED. Finally, from high-dose implantation, EOR defects are modeled and we derived a formula to describe the time-dependence for Ostwald ripening of EOR defects, which is more significant at higher temperatures and longer annealing times. The simulation satisfactorily predicts the TED for annealing conditions, where the calculations overestimate the diffusion without taking Ostwald ripening into account.

Effects of glucocorticoid treatment and acute passive immunization with growth hormone-releasing hormone and somatostatin antibodies on endogenous and stimulated growth hormone secretion in the male rat

1991 ◽  
Vol 131 (1) ◽  
pp. 75-86 ◽  
Author(s):  
J. Miell ◽  
R. Corder ◽  
P. J. Miell ◽  
C. McClean ◽  
R. C. Gaillard
Keyword(s):  
Growth Hormone ◽  
Low Dose ◽  
Growth Hormone Secretion ◽  
Blood Sampling ◽  
Male Rat ◽  
High Dose ◽  
Glucocorticoid Treatment ◽  
Releasing Hormone ◽  
Gh Secretion ◽  
Medium Dose

ABSTRACT Despite causing marked inhibition of somatic growth, glucocorticoids enhance both the response to GH-releasing hormone (GHRH) and the amplitude of naturally occurring GH secretory pulses in the male rat. The relative contribution of the two major hypothalamic regulatory factors for GH (somatostatin and GHRH) to these observed effects remains speculative. In the present studies, we have investigated endogenous and stimulated GH release in rats pretreated with glucocorticoid or vehicle, and the effects of passive immunoneutralization of somatostatin or GHRH. In an initial study, four groups of eight rats were treated with either saline or various doses of a depot preparation of betamethasone: low dose, 0·85 mg; medium dose, 1·7 mg; high dose, 3·4 mg. All doses significantly suppressed body weight gain, total adrenal weight and concentrations of both plasma corticosterone and pituitary ACTH. Seven days after betamethasone treatment, GH responses to an i.v. injection of 1 μg human GHRH(1–29) were evaluated during pentobarbitone anaesthesia. Compared with saline-treated controls (peak GH concentration of 506·0±68·5 μg/l), peak GH levels were enhanced by the low dose (704·4±47·8 μg/l, P<0·05), unaltered by the medium dose (543±65·8 μg/l) and suppressed by the high dose (312·7±55·2 μg/l, P<0·05) of betamethasone. Similarly, the area under the secretory curves was increased by 46% following the low dose (P<0·01), unaltered by the medium dose and reduced by 33% after the high dose of betamethasone. In a second study, rats were pretreated for 7 days before blood sampling with either the medium dose of betamethasone or saline. On day 5, 48 h before blood sampling, an indwelling venous catheter was fitted enabling sampling of conscious rats. On the day of study, blood samples were taken at 30-min intervals over an initial 2-h period (10.00–12.00 h). Following the sample at 12.00 h, rats were given the reconstituted and dialysed immunoglobulin fraction from either control sheep serum (NSIgG), sheep anti-rat GHRH serum (GHRHab) or sheep anti-somatostatin serum (SRIHab), and samples were taken for a further 90 min (12.30–14.00 h). Directly after the sample at 14.00 h, GH stimulation was effected in all rats using 1 μg human GHRH(1–29) with samples taken at 5, 10, 20 and 40 min following stimulation. During the initial sampling period, mean GH levels were significantly (P<0·005) higher in steroidpretreated animals than in saline-pretreated controls (29·3±5·8 vs 13·2±1·6 μg/l), with a higher amplitude secretory pulse occurring at 11.30 h (80·7±18·6 vs 26·4±4·1 μg/l, P < 0·01). Administration of GHRHab to saline-pretreated animals did not alter mean GH levels when compared with animals receiving control NSIgG (saline plus NSIgG, 9·3±1·1; saline plus GHRHab, 8±1·1 μg/l, P = NS). In contrast, the raised mean GH levels seen in betamethasone-pretreated rats receiving NSIgG (12·3 ±1·1 μg/l) were reduced by GHRHab administration (7·6±1·1 μg/l); these levels were not different from those of the saline-pretreated group suggesting that the observed permissive effect of glucocorticoids on GH secretion is mediated through enhanced GHRH activity. SRIHab increased mean basal GH levels to a similar extent in both saline- and betamethasone-pretreated groups (17·4±1·2 μg/l and 19·3 ±1·1 μg/l respectively, P<0·01 vs comparable NSIgG group). Administration of the various immunoglobulin fractions had no effect on GHRH-stimulated GH secretion except when SRIHab was given to betamethasone-pretreated animals, resulting in a significantly increased peak response (1467±93 μg GH/l, P<0·001) when compared with either saline- or betamethasone-pretreated rats given NSIgG (643±95 and 791±92 μg/l respectively). This enhancement following SRIHab administration was not seen in saline-pretreated animals (893±180 μg GH/l). These results imply that glucocorticoid treatment increases basal GH levels through a GHRH-dependent mechanism and also increases pituitary sensitivity to exogenous GHRH when inhibitory somatostatin tone is blocked. Journal of Endocrinology (1991) 131, 75–86

Fluid balance, electrolyte profiles and plasma parathyroid hormone concentrations in ewes treated with epidermal growth factor

1992 ◽  
Vol 135 (1) ◽  
pp. 91-101 ◽  
Author(s):  
C. B. Gow ◽  
M. Wilkinson ◽  
M. J. Silvapulle ◽  
G. P. M. Moore
Keyword(s):  
Parathyroid Hormone ◽  
Growth Factor ◽  
Fluid Balance ◽  
Low Dose ◽  
Plasma Concentrations ◽  
Urine Volume ◽  
High Dose ◽  
Epidermal Growth ◽  
Higher Doses ◽  
Medium Dose

ABSTRACT The infusion of low doses of epidermal growth factor (EGF) into lactating ewes stimulates water intake and urine volume. The plasma concentrations and daily output of various electrolytes in milk and urine are also affected. We have investigated this further by recording the effects of EGF infusion on fluid balance, electrolyte profiles and plasma concentrations of glucose and parathyroid hormone (PTH) in non-pregnant, non-lactating ewes. Twenty-four animals (n= 8 per group) received infusions of 100 ml saline/day into the jugular vein for 10 days (days 1–10) followed by EGF at a dose rate of either 1 (low dose), 5 (medium dose) or 10 (high dose) μg/kg liveweight per day in 100 ml saline for 5 days (days 11–15). All ewes then received an infusion of 100 ml saline/day for 10 days (days 16–25). Most plasma and urine samples had undetectable concentrations of EGF-immunoreactive material during the periods of saline infusion. During EGF infusion, the highest amounts of EGF infusate excreted in urine were 1·6, 5·9 and 5·6% for ewes in low, medium and high dose groups respectively. Water intake increased by 17% (0·5 kg), 88% (2·5 kg) and 89% (2·3 kg) and urine volume increased by 29% (0·5 kg), 108% (2·2 kg) and 134% (2·1 kg) for the three groups respectively. Fluid balance and feed intake were not affected by EGF infusion, but the output of faecal dry matter was reduced in ewes receiving the two higher doses of EGF. All levels of EGF resulted in hypocalcaemia, increased plasma PTH concentrations and hypermagnesaemia. There was no effect of EGF on plasma concentrations of K+ and glucose or on daily urinary excretion of K+ and Mg2+. The only response to the low dose was a reduced plasma concentration of Na+ and an increased daily urinary urate excretion. The two higher doses increased the daily urinary excretion of Na+, PO43− and urate, but had no effect on the respective concentrations in plasma. Urinary Ca2+ excretion was reduced only during infusion of the medium dose of EGF. The responses of most variables were similar during infusion of the medium and high doses of EGF. All three doses of EGF induced polydipsic and diuretic responses in ewes, and infusions of 5–10 μg EGF/kg liveweight per day affected renal excretion of Ca2+, Na+ and PO3−4. We interpret the responses of the kidney and plasma PTH concentrations as a means of maintaining the homeostasis of plasma profiles of electrolytes. Journal of Endocrinology (1992) 135, 91–101

scholarly journals Treatment-related damage in elderly-onset ANCA associated vasculitis: Safety outcome analysis of two nationwide prospective cohort studies

2020 ◽  
Author(s):  
Ken-ei Sada ◽  
Keiji Ohashi ◽  
Yosuke Asano ◽  
Keigo Hayashi ◽  
Michiko Morishita ◽  
...  
Keyword(s):  
Cohort Studies ◽  
Low Dose ◽  
Damage Index ◽  
Antineutrophil Cytoplasmic Antibody ◽  
Outcome Analysis ◽  
High Dose ◽  
Rapid Reduction ◽  
Anca Associated Vasculitis ◽  
The Mean ◽  
Medium Dose

Abstract Background: It is not elucidated that treatment-related damage in elderly patients with antineutrophil cytoplasmic antibody (ANCA)–associated vasculitis (AAV). Methods: Elderly (≥75 years of age) patients were enrolled from two nationwide prospective inception cohort studies. The primary outcome was 12-month treatment-related Vasculitis Damage Index (VDI) score. Secondary outcomes included serious infections within 6 months, total VDI score, remission, and relapse. Patient characteristics and outcomes were compared across three different initial glucocorticoid (GC) dose groups: high-dose, prednisolone (PSL) ≥ 0.8 mg/kg/day; medium-dose, 0.6 ≤ PSL < 0.8 mg/kg/day; and low-dose, PSL < 0.6 mg/kg/day. Results: Of the 179 eligible patients, the mean age was 80.0 years; 111 (62%) were female. The mean Birmingham Vasculitis Activity Score was 16.1. Myeloperoxidase-ANCA findings were positive in 168 (94%) patients, while proteinase3-ANCA findings were positive in 11 (6%). The low-dose group was older and had higher serum creatinine levels than the other groups. There were no statistically significant intergroup differences in remission or relapse, whereas serious infection developed more frequently in the high-dose (29 patients [43%]) than the low-dose (13 patients [22%]) or medium-dose (10 patients [19%]) groups (p = 0.0007). Frequent VDI items at 12 months included hypertension (19%), diabetes (13%), atrophy and weakness (13%), osteoporosis (8%), and cataracts (8%). Logistic regression analysis revealed that GC dose at 12 months (odds ratio, 1.14; 95% confidence interval, 1.00–1.35) was a predictor for diabetes.Conclusion: A reduced initial GC dose with rapid reduction might be required to ensure the safe treatment of elderly AAV patients.

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