PGF2 Research Articles

Noradrenaline and adrenoreceptors regulate prostaglandin F2A formation in endometrium after experimentally-induced inf lammation in the pig

Abstract Endometritis is the most common pathology in animals. However, in the context of an inflamed endometrium, alterations occur in the production of prostaglandins (PG s) and the noradrenergic innervation of the uterus, although the precise relationship between noradrenaline (NA), adrenoreceptors (AR s), and the output of PG F2α remains unclear. To clarify this issue, the participation of α1-, α2- and β-AR s in NA-influenced PG F synthase (PG FS) and PG 9-ketoreductase/carbonyl reductase (CBR1) protein abundances in the porcine inflamed endometrium, and the secretion of PG F2α from the tissue were determined. E. coli suspension (E. coli group) or saline (CO N group) was injected into the uterine horns. After eight days, severe acute endometritis was diagnosed in the E. coli group. Endometrial explants were treated with NA and/or α1-, α2- and β-AR s antagonists. In the CO N and E. coli groups, NA increased endometrial PG FS and CBR1 protein abundances and PG F2α secretion, compared to the control values (obtained from an endometrium that had not undergone any in vitro treatment). In the E. coli group, NA-stimulated CBR1 protein abundance and PG F2α release were higher, while PG FS protein abundance was lower than in the CO N group. In the latter group, the antagonists of α1A-, α1D-, α2B- and α2C-AR s isoforms and β2- AR s subtype decreased NA-stimulated PG FS protein abundances, compared to NA action alone. In the E. coli group, this effect on PG FS abundances evoked α1D-, α2C-, β1- and β2-AR s antagonists with NA. Antagonists of α1B-, α2B-, β1- and β2-AR s in the CO N group and antagonists of α1B-, α1D-, α2A-, α2C-, β1- and β2-AR s in the E. coli group eliminated a rise in the NA-stimulated CBR1 abundance of protein versus the NA influence alone. In comparison to NA effect alone, α1D-, α2C- and β2-AR s antagonists with NA reduced PG F2α secretion in both the CON and E. coli groups. Such effect on PG F2α release was also exerted in the E. coli group by α1B-, α2A- and β1-AR s antagonists with NA. Summarizing, in the porcine inflamed endometrium, NA increases PG FS protein abundance via α1D-, α2C- and β(1, 2)-AR s, and CBR1 protein abundance and PG F2α release by α1(B, D)-, α2(A, C) and β(1, 2)-AR s. The obtained findings suggest that, in an indirect manner, NA may affect the PG F2α-regulated processes by influencing its production and secretion. The results could offer new targets for drugs to regulate inflammation and improve uterine and ovarian functions.

Primary dysmenorrhea: pathophysiology, improvement of diagnostic and treatment possibilities

Primary dysmenorrhea is one of the types of gynecological pathology, the frequency of which is 45-53% in girls and young women aged 13 to 45 years, which is not given enough attention in diagnosis and treatment.The pathogenesis takes into account the increased secretion of prostaglandins F2α and E2 in the period from the beginning of pregnancy to birth, which cause pelvic pain.The diagnosis is based on the patient's history, symptoms, examination, the treatment is aimed at improving the quality of life by prescribing nonsteroidal anti-inflammatory drugs, hormonal contraceptives and the use of non-medicinal means. Purpose: to specify some links in the pathogenesis of primary dysmenorrhea and, taking into account its multicomponent nature, to evaluate the effectiveness of the proposed treatment. Under observation there were 68 women who were divided into 2 groups: 34 women with primary dysme­norrhea (main group) and 34 healthy women (control group).The diagnosis of primary dysmenorrhea was established on the basis of complaints of patients about painful menstruation and accompanying symptoms during consultations with a gynecologist and endocrinologist.Treatment of patients in the main group was carried out with the drug, which includes Vitex agnus castus L, indinol-3-carbinol, 3,3-diindolyl-methane, extract of passionflower, California Eschscholzia. In women with typical symptoms, dysmenorrhea can be diagnosed on the basis of medical information without physical pelvic examination, empirical treatment should be initiated, including nonsteroidal anti-inflammatory drugs and/or oral contraceptives, because it is a functional condition resulting from an imbalance in the hormonal regulation of myometrial contraction. As a result of the proposed treatment, pain disappeared completely in 70% of patients (23/34), as well as vegetative-vascular (from 18 to 2% of patients), vegetative (from 10 to 1% of patients), metabolic and endocrine disorders (from 15 to 2% female patients), disorders of the emotional and mental sphere (from 25 to 10% of patients). Improvement in quality of life was noted by 80% (26/34) of patients, improvement in working capacity – by 70% (23/34) of patients, after 2 months – by 95% (31/34) and 85% (28/34), respectively, after 3 months all examined patients with primary dysmenorrhea noted an improvement in the quality of life and an increase in work capacity. Sexually active women with symptoms of inflammatory processes of the genital tract, severe dysmenorrhea should undergo pelvic examination. In view of safety and high therapeutic efficiency, the drug based on Vitex agnus castus can be offered for the treatment over the period lasting at least for3 months for girls and young women aged 13 to 45 years with primary dysmenorrhea.

Spanlastic-laden nanogel as a plausible platform for dermal delivery of bimatoprost with superior cutaneous deposition and hair regrowth efficiency in androgenic alopecia

Bimatoprost (BIM) is a prostaglandin F2α analogs originally approved for the treatment of glaucoma and ocular hypertension. Recent studies have highlighted its potential to boost hair growth. The objective of this investigation is to challenge the potential of spanlastics (SLs) as a surfactant-based vesicular system for promoting the cutaneous delivery of BIM for the management of alopecia. BIM-loaded spanlastics (BIM-SLs), composed of Span as the main vesicle component and Tween as the edge activator, were fabricated by ethanol injection method. The formulated BIM-SLs were optimized by 23 full factorial design. The optimized formula (F1) was characterized for entrapment efficiency, surface charge, vesicle size, and drug release after 12 h (Q12h). The optimized formula (F1) exhibited high drug entrapment efficiency (83.1 ± 2.1%), appropriate zeta potential (−19.9 ± 2.1 mV), Q12h of 71.3 ± 5.3%, and a vesicle size of 364.2 ± 15.8 nm, which favored their cutaneous accumulation. In addition, ex-vivo skin deposition studies revealed that entrapping BIM within spanlastic-based nanogel (BIM-SLG) augmented the dermal deposition of BIM, compared to naïve BIM gel. Furthermore, in vivo studies verified the efficacy of spanlastic vesicles to boost the cutaneous accumulation of BIM compared to naive BIM gel; the AUC0-12h of BIM-SLG was 888.05 ± 72.31 μg/mL.h, which was twice as high as that of naïve BIM gel (AUC0-12h 382.86 ± 41.12 μg/mL.h). Intriguingly, BIM-SLG outperforms both naïve BIM gel and commercial minoxidil formulations in stimulating hair regrowth in an androgenetic alopecia mouse model. Collectively, spanlastic vesicles might be a potential platform for promoting the dermal delivery of BIM in managing alopecia.

A concise and scalable chemoenzymatic synthesis of prostaglandins

Prostaglandins have garnered significant attention from synthetic chemists due to their exceptional biological activities. In this report, we present a concise chemoenzymatic synthesis method for several representative prostaglandins, achieved in 5 to 7 steps. Notably, the common intermediate bromohydrin, a radical equivalent of Corey lactone, is chemoenzymatically synthesized in only two steps, which allows us to complete the synthesis of prostaglandin F2α in five steps on a 10-gram scale. The chiral cyclopentane core is introduced with high enantioselectivity, while the lipid chains are sequentially incorporated through a cost-effective process involving bromohydrin formation, nickel-catalyzed cross-couplings, and Wittig reactions. This cost-efficient synthesis route for prostaglandins holds the potential to make prostaglandin-related drugs more affordable and facilitate easier access to their analogues.

Can Non-invasive Hormone Profiling in Female Tigers (Panthera tigris) could Differentiate True Pregnancy and Pseudopregnancy

Background: The tigers (Panthera tigris) being the world’s largest cat are one of the critically endangered species which are at the verge of extinction. The pseudopregnancy or false pregnancy or pseudocyesis is a syndrome which is characterized by the presence of signs of pregnancy in a non-pregnant animal. False lactation or nervous lactation are the terms which are used to refer pseudopregnancy as there is very low quantities of milk production is seen. This condition can be differentiated with true pregnancy by using hormones and by correlation of behavioral symptoms of pregnancy with hormonal profiles for confirmation of pregnancy in female tigers. Methods: Total seven animals were used in the experiment and was divided into three groups (G I, G II and G III). Group G I consist of 03 non-bred animals of less than 9 years of age, G II consisted of 03 non-bred animals of more than 9 years of age and G III was consisted of 03 animals after mating irrespective of their age. Faecal samples were collected non-invasively from enclosures weekly up to 14 weeks. Only fresh samples (deposited within 24 h) was chosen, urine puddles were aspirated from the floor. The extracted samples were then analysed for estimation of relaxin, progesterone and prostaglandin F2 alpha metabolite (PGFM) by enzyme linked immunosorbent assay (ELISA). Result: Relaxin the hormone for early detection of pregnancy in felines gives elevated concentration from 49 days after mating and attains a peak during parturition, whereas, no elevation or peak was observed in pseudopregnant animals. Progesterone, in true pregnancy remains elevated from 7th week onwards after mating, it gets dropped after 7th week with wide fluctuation in pseudopregnant animals. PGFM elevation observed after nine weeks of gestation in true pregnancy whereas, no elevation will be observed in pseudopregnant tigress.

Effect of PTFGRN Expression on the Proteomic Profile of A431 Cells and Determination of the PTGFRN Interactome.

Prostaglandin F2 receptor negative regulator (PTGFRN) is a transmembrane protein whose expression has been previously implicated in cancer metastasis. However, the exact molecular mechanisms by which PTGFRN influences cancer progression are still unknown. As such, our laboratory set out to investigate how PTGFRN knockdown affected the expression of other proteins. We also carried out coimmunoprecipitation experiments using a monoclonal anti-PTGFRN antibody. We employed mass spectrometry-based proteomics for both experiments to identify proteins that were associated with PTGFRN. Our data show that PTGFRN knockdown increased pathways related to innate immune responses and decreased pathways associated with the synthesis of metabolic precursors and protein processing, among others. Additionally, the coimmunoprecipitation experiments indicated that PTGFRN is associated with proteins involved in processing and metabolism, as well as VEGF signaling molecules. These results highlight the role of PTGFRN as a protein processing regulator, which may be influencing cancer progression.

AKR1C3 in carcinomas: from multifaceted roles to therapeutic strategies.

Aldo-Keto Reductase Family 1 Member C3 (AKR1C3), also known as type 5 17β-hydroxysteroid dehydrogenase (17β-HSD5) or prostaglandin F (PGF) synthase, functions as a pivotal enzyme in androgen biosynthesis. It catalyzes the conversion of weak androgens, estrone (a weak estrogen), and PGD2 into potent androgens (testosterone and 5α-dihydrotestosterone), 17β-estradiol (a potent estrogen), and 11β-PGF2α, respectively. Elevated levels of AKR1C3 activate androgen receptor (AR) signaling pathway, contributing to tumor recurrence and imparting resistance to cancer therapies. The overexpression of AKR1C3 serves as an oncogenic factor, promoting carcinoma cell proliferation, invasion, and metastasis, and is correlated with unfavorable prognosis and overall survival in carcinoma patients. Inhibiting AKR1C3 has demonstrated potent efficacy in suppressing tumor progression and overcoming treatment resistance. As a result, the development and design of AKR1C3 inhibitors have garnered increasing interest among researchers, with significant progress witnessed in recent years. Novel AKR1C3 inhibitors, including natural products and analogues of existing drugs designed based on their structures and frameworks, continue to be discovered and developed in laboratories worldwide. The AKR1C3 enzyme has emerged as a key player in carcinoma progression and therapeutic resistance, posing challenges in cancer treatment. This review aims to provide a comprehensive analysis of AKR1C3's role in carcinoma development, its implications in therapeutic resistance, and recent advancements in the development of AKR1C3 inhibitors for tumor therapies.

Urinary prostanoids are elevated by anti-TNF and anti-IL6 receptor disease-modifying antirheumatic drugs but are not predictive of response to treatment in early rheumatoid arthritis

BackgroundDisease-modifying antirheumatic drugs (DMARDs) are widely used for treating rheumatoid arthritis (RA). However, there are no established biomarkers to predict a patient’s response to these therapies. Prostanoids, encompassing prostaglandins, prostacyclins, and thromboxanes, are potent lipid mediators implicated in RA progression. Nevertheless, the influence of DMARDs on prostanoid biosynthesis in RA patients remains poorly understood. This study aims to assess the impact of various DMARDs on urinary prostanoids levels and to explore whether urinary prostanoid profiles correlate with disease activity or response to therapy.MethodsThis study included 152 Swedish female patients with early RA, all rheumatoid factor (RF) positive, enrolled in the NORD-STAR trial (registration number: NCT01491815). Participants were randomized into four therapeutic regimes: methotrexate (MTX) combined with (i) prednisolone (arm ACT), (ii) TNF-α blocker certolizumab pegol (arm CZP), (iii) CTLA-4Ig abatacept (arm ABA), or (iv) IL-6R blocker tocilizumab (arm TCZ). Urine samples, collected before start of treatment and at 24 weeks post-treatment, were analyzed for tetranor-prostaglandin E metabolite (tPGEM), tetranor-prostaglandin D metabolite (tPGDM), 2,3-dinor thromboxane B2 (TXBM), 2,3-dinor-6-keto prostaglandin F1a (PGIM), leukotriene E4 (LTE4) and 12-hydroxyeicosatetraenoic acid (12-HETE) using liquid chromatography–mass spectrometry (LC–MS). Generalized estimating equation (GEE) models were used to analyze the change in urinary eicosanoids and their correlations to clinical outcomes.ResultsPatients receiving MTX combined with CZP or TCZ exhibited significant elevations in urinary tPGEM and TXBM levels after 24 weeks of treatment. Other eicosanoids did not show significant alterations in response to any treatment. Baseline urinary eicosanoid levels did not correlate with baseline clinical disease activity index (CDAI) levels, nor with changes in CDAI from baseline to week 24. Their levels were also similar between patients who achieved CDAI remission and those with active disease at week 24.ConclusionsTreatment with anti-TNF or anti-IL6R agents in early RA patients leads to an increased systemic production of proinflammatory and prothrombotic prostanoids. However, urinary eicosanoid levels do not appear to be predictive of the response to DMARDs therapy.

(007) Impact of Non-Invasive Oestrus Synchronization on Sexual Behavior in Intact Sprague Dawley Rats

Abstract Introduction Female sexual behavior animal translation data do not translate to acute results and neither current synchronization invasive models (ovariectomy+Estrogen “E2”+Progesterone “P4”) guarantee hormonal physiological values. Objective Therefore, our study investigated the effect of non-invasive oestrus synchronization on sexual behavior. Methods Methods: Twelve week old Sprague Dawley intact rats were synchronized and randomly divided into three groups each of n=8-10. Each group received a dose of prostaglandin F2α “PGF2α” (75, 125 and 250 μg) at days 0 and 3, and 1 mg of progesterone at day four. Vaginal cytology was performed to determine the point of the cycle and the animals underwent an incentive test. To investigate the sexual behavior, the synchronized female rats were paired with trained male rats, and behavior recorded. The videos were quantified by trained operators. Sexual behavior was quantified as paracopulatory (hops, darts, ear wiggling, sniffing from male and from female) or copulatory (male mounting female, thrusting from male, intromission, ejaculation, female for sniffing position, lordosis, orgasm, female mounting on male, female rotation in her axis and female licking itself), E2 and Follicle Stimulating hormone (FSH) in plasma and E2 in vaginal fluid were quantified by immune assay. Results Based on the vaginal cytology, the proportion of animals in proestrus (p), estrus (e) or metestrus (m) was calculated with values of 0.5 (p), 0.25 (e) and 0.25 (m) for the group which received 0.75 μg PGF2α; 0.4 (p), and 0.6 (e) for the group which received 0.125 μg PGF2α; 0.2 (p), 0.5 (e), and 0.3 (m) for the group which received 250 μg PGF2α. The numbers of hops was higher in the proestrus compared to the metestrus, and of mounts higher in the estrus compared to the metestrus phase. The sexual encounter profile and E2 levels were unchanged with the different PGF2α doses, but in the high-dose group no hops were observed. The levels of sexual behavior was comparable to previous studies of hormone substituted ovariectomized rats, but E2 and FSH plasma levels were in a physiological range. Conclusions In summary, the non-invasive synchronization protocol could substitute the state-of-the-art used ovariectomized+hormone (E2) model. Our findings suggest the largest part of rats were synchronized in the oestrus phase with 125 μg PGF2α. but the 250 μg had also a positive profile. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Initiator Pharma A/S.

15 Influence of early progesterone removal on follicular development, estrus expression, and pregnancy rates in presynchronized beef cows

Abstract The objective of this study was to evaluate the impact of early progesterone removal on pregnancy rates to fixed-time artificial insemination (FTAI) in presynchronized beef cows. Postpartum beef cows (n = 922; body condition score = 4.9 ± 0.77; days postpartum = 77.3 ± 20.77 d) from 7 locations were enrolled in a completely randomized design. Within location, cows were stratified by parity and days postpartum before being randomly assigned to 1 of 2 treatments: 1) 7&7 Synch: Cows received a controlled internal drug release insert (CIDR) and a 25-mg injection of prostaglandin F2α (PG1; d 0). Cows received 100 μg of GnRH on d 7 and the CIDR was removed 7 d later, concurrently with a second prostaglandin F2α (PG2) injection (d 14). Cows received a second injection of GnRH and were FTAI 60-66 h after PG2 (d 17); 2) 7&6 Synch: Cows received the same treatment as 7&7 Synch cows; however, CIDR removal and PG2 were performed on d 13, and cows were FTAI 60-66 h after CIDR removal. Ovarian ultrasonography was performed to determine follicle diameter and CL presence at PG2 and FTAI in a subset of cows (n = 40). Estrus detection patches were utilized to evaluate estrus expression between PG2 and FTAI. Pregnancy diagnosis was performed 28-35 d after FTAI. Cows exposed to the 7&7 Synch tended to have larger follicle diameter at PG2 compared with 7&6 Synch cows (P = 0.09); however, there were no differences in follicle diameter at FTAI. Cows that expressed estrus tended to have larger follicle diameter at FTAI compared with cows that failed to express estrus (P = 0.10). Follicular growth between PG2 and FTAI, percentage of cows with a corpus luteum at PG2, and ovulation rate were not impacted by treatment (P ≥ 0.56). No differences were determined between treatments for estrus expression (7&7 Synch: 77.1 ± 5.91; 7&6 Synch: 74.6 ± 6.28; P = 0.50) or pregnancy rates to FTAI (7&7 Synch: 60.1 ± 4.32; 7&6 Synch: 58.2 ± 4.26; P = 0.69). In conclusion, early removal of progesterone did not impact pregnancy rates in presynchronized beef cows. Thus, the 7&6 Synch protocol described herein may be used as an alternative to the conventional 7&7 Synch protocol without compromising fertility.

12 Impact of progesterone signaling on tumor necrosis factor-alpha synthesis by luteal macrophages

Abstract Understanding the mechanism in which luteolysis occurs is an important step in improving the reproduction of livestock in the industry. The corpus luteum (CL) is a transient endocrine organ found on the ovary responsible for progesterone secretion. It is well known that prostaglandin F2-alpha (PGF) is responsible for luteal regression, however, the process by which luteal cells (LC) become sensitive to the luteolytic effects of PGF remain unclear. Therefore, the objective of this study was to determine if suppression of progesterone (P4) synthesis or blocking P4 signaling activates tumor necrosis factor (TNF)-alpha production by the luteal macrophages. Ovaries were collected from an abattoir and transported to the laboratory on ice. Ovary status (i.e. approximate days post ovulation), was determined by phenotypic description of ovarian structures. Only ovaries (n = 5) containing midcycle (d 12/13 post ovulation) CL were used in these experiments. Luteal tissue was isolated and dissociated to collect LC and and luteal macrophages (MAC). Isolated cells were incubated independently at 37°C at 5% CO2 for 24 h in 24-well plates. Luteal cells and MACs were then harvested and placed in the following conditions: 1) LC + MAC + DMSO (control), 2) LC + MAC + trilostane (TRIL 1.0mg/mL), 3) LC + MAC + TRIL 1.5 mg/mL, 4) MACS only, 5) MACS +LPS. Supernatant was harvested at 24 h whereas both cells and supernatant were harvested after 48 h, in vitro. Supernatant was immediately frozen for subsequent P4 analysis, via RIA. Data results were analyzed using a MIXED procedure of SAS and evaluated the effects of cell-type, treatment, and culture time. Statistical significance was determined at P < 0.05 and a tendency at 0.05 < P < 0.10. Overall, TRIL treatment reduced supernatant P4 concentration (8.3 and 6.8 ng/mL to 1.0 and 1.5 mg/mL TRIL, respectively) compared with controls (22.8 ng/mL; P = 0.0012); however there was not a difference when comparing the TRIL treated cultures. Progesterone concentrations tended to decrease over time (P = 0.0661). At 24 h in vitro, P4 concentrations were suppressed in 1.0 mg/mL TRIL and 1.5 mg/mL TRIL treated wells (5.3 and 4.2 ng/mL, respectively), compared with controls (16.4 ng/mL; P = 0.0014). Whereas P4 concentrations were reduced in the 1.5 mg/mL TRIL treated wells (1.6 ng/mL) compared with controls (9.95 ng/mL; P = 0.0419) and tended to differ in the1.0 mg/mL TRIL treated wells (2.4 ng/mL; P = 0.0634), at 48 h in vitro. Progesterone concentrations were undetectable in wells containing MAC only. Collectively, TRIL treatment effectively reduced P4 synthesis in LC cultures. Therefore, the greater concentration of TRIL is needed for P4 suppression in longer term culture systems. Additional analyses will be conducted to determine the effect of P4 suppression on macrophage function.

Improving the efficiency of estrus synchronization in cows.

This study aimed to determine the effects of biologically active substances and electrical stimulation of the uterus in cows on the effectiveness of estrus synchronization. Ninety (n = 90) Kazakh white-headed cows were synchronized with two injections of gonadotropin-releasing hormone on days 0 and 9 and prostaglandin F2α on day 7. The cows were divided into six groups and, during the protocol, treated with biologically active substances (Tetramag, Selevetum, antiseptic-stimulator Dorogov 2 fraction, groups 2, 3, and 4). Cows in groups 5 and 6 were treated with the same substances but additionally had electrical stimulation of the uterus, while cows in group 1 were left untreated and served as a control. The results have shown that on Day 0, no differences were observed in E2 concentrations between the groups. However, on the 10th day, a significant disparity was noted in the E2 level among cows in group 6 compared to groups 2, 3, 4, and the control group. Conversely, no significant differences were observed between groups 5 and 6. Likewise, the fertility rate in cows from group 6 was significantly higher compared to groups 2, 3, 4, and the control group, with no significant differences between groups 5 and 6. It can be concluded that the utilization of electrical stimulation of the uterus and the inclusion of certain biological substances during the estrus synchronization protocol demonstrate a positive effect on the reproductive performance of beef cattle in Kazakhstan.

Absence of Systemic Oxidative Stress and Increased CSF Prostaglandin F2α in Progressive MS.

Absence of Systemic Oxidative Stress and Increased CSF Prostaglandin F2α in Progressive MS.

Ginseng fermentation solution affects the gut microbiota in zebrafish with alcoholic liver disease via PI3K/Akt pathway

BackgroundGinsenosides have received increased amounts of attention due to their ability to modulate the intestinal flora, which may subsequently alleviate alcoholic liver disease (ALD). The effects of ginseng fermentation solution (GFS) on the gut microbiota and metabolism in ALD patients have not been explored. PurposeThis research aimed to explore the regulatory effect of GFS on ALD both in vitro and in vivo. MethodThis study assessed the anti-ALD efficacy of GFS using an LO2 cell model and a zebrafish model. Untargeted metabolomics was used for differentially abundant metabolite analysis, and high-throughput 16S rRNA sequencing was used to examine the effect of GFS on ALD. ResultsThe LO2 cell line experiments demonstrated that GFS effectively mitigated alcohol-induced oxidative stress and reduced apoptosis by upregulating PI3K and Bcl-2 expression and decreasing the levels of malondialdehyde, total cholesterol, and triglycerides. In zebrafish, GFS improved morphological and physiological parameters and diminished oxidative stress-induced ALD. Meanwhile, the results from Western blotting indicated that GFS enhanced the expression of PI3K, Akt, and Bcl-2 proteins while reducing Bax protein expression, thereby ameliorating the ALD model in zebrafish. Metabolomics data revealed significant changes in a total of 46 potential biomarkers. Among them, metabolites such as prostaglandin F2 alpha belong to arachidonic acid metabolism. In addition, GFS also partly reversed the imbalance of gut microbiota composition caused by alcohol. At the genus level, alcohol consumption elevated the presence of Flectobacillus, Curvibacter, among others, and diminished Elizabethkingia within the intestinal microbes of zebrafish. Conversely, GFS reversed these effects, notably enhancing the abundance of Proteobacteria and Archaea. Correlation analyses further indicated a significant negative correlation between prostaglandin F2 alpha, 11,14,15-THETA, Taurocholic acid and Curvibacter. ConclusionThis study highlights a novel mechanism by which GFS modulates anti-ALD activity through the PI3K/Akt signalling pathway by influencing the intestinal flora-metabolite axis. These results indicate the potential of GFS as a functional food for ALD treatment via modulation of the gut flora.

Oestrus induction and fertility response in post-partum anoestrus dairy cows treated with homeopathic medicine

The study was carried out to evaluate the effect of two homeopathic medicines to induce oestrus in 36 postpartum anoestrus crossbred Jersey dairy cows. Group 1 cows were fed a mineral mixture, and group 2 and 3 were medicated with Pulsatilla and Sepia drenched at the dose rate of 20 mL twice daily for 8 days, respectively. The mean plasma progesterone level (ng/mL) estimated using a radio immunoassay kit was significantly higher in group 2, followed by group 3 before and after medication on day 28 and 60, post artificial insemination (AI). Out of 12 induced for oestrus, 10 cows (83.33 %) and 9 cows (75.0 %) in group 2 and 3 were confirmed pregnant by rectal palpation on day 60 post AI, respectively. Both the homeopathic medicines showed 100% oestrus induction with a mean interval of 25.17 to 25.50 days, and Pulsatilla medicated cows showed improved fertility response than Sepia. The significantly higher level of progesterone after the withdrawal of homeopathic medication might be due to the release of circulating hormone, which blocks estradiol negative feedback. The subsequent increase reduces uterine PGF2 alpha secretion for a better conception rate.

Estrous characteristics and steroid concentrations in Aceh cows (Bos indicus) after estrus synchronization using prostaglandin F2a

Abstract. Hafizuddin, Husnurrizal, Thasmi CN, Herrialfian, Salsabil NL, Siregar TN, Sutriana A. 2024. Estrous characteristics and steroid concentrations in Aceh cows (Bos indicus) after estrus synchronization using prostaglandin F2a. Biodiversitas 25: 322-328. This study aims to determine the estrous characteristics and steroid concentrations during estrus in Aceh cattle after estrus synchronization using prostaglandin F2a. This study involved 9 Aceh cows with good reproductive status, not pregnant, and has shown at least 2 regular cycles, which were divided into 3 treatment groups. The first group (GI) received a single injection treatment, the second group (G2) received a double injection treatment with an interval of 10 days, and the third group (G3) was also treatment double injection with an interval of 12 days. Symptoms and responses to estrus were reported descriptively, and the data collected on onset, intensity, and duration of estrus were analyzed using one-way analysis of variance followed by Duncan's test. The results showed that estrus symptoms were almost similar and estrus response was 100% in all three treatment groups. The onset of estrus in G1, G2, and G3 did not differ significantly (P>0.05) with values of 68.00±18.33, 60.00±0.00, and 72.00±24.00 hours, respectively. The intensity of estrus in G1, G2, and G3 showed significant differences (P<0.05) with scores of 3.00±0.00, 5.00±0.00 and 4.33±1.15, respectively. The duration of estrus showed no significant difference in G1, G2, and G3 (P>0.05) with durations of 32.00±18.33, 72.00±0.00, and 52.00±38.57 hours, respectively. The concentrations of estradiol and progesterone obtained also showed no significant differences (P>0.05) in G1, G2, and G3, with respective concentrations of 91.67±24.77, 132.00±23.15, 89.47±24.19 pg/mL and 0.98±0.59, 0.86±0.18, 1.00±0.17 ng/mL. In conclusion, estrus synchronization with PGF2a single injection and double injection in Aceh cattle was found to have 100% estrus response. However, PGF2a with double injection showed higher estrus intensity compared to single injection. In Aceh cows, the recommended protocol for estrus synchronization using PGF2a is double injection.

Intrauterine devices influence prostaglandin secretion by equine uterus: in vitro and in vivo studies

BackgroundIntrauterine devices (IUD) are used in the veterinary practice as the non-pharmacological method of oestrus suppression in mares. When placed in the uterus, IUD create a physical contact with the endometrium that mimics the presence of an equine embryo. However, the mechanism of their action has not been fully elucidated. The objective of the present study was to examine the effect of mechanical stimulation of IUD on mare`s endometrium in both in vitro and in vivo study. For this purpose, we demonstrated the effect of IUD on prostaglandin (PG) F2α and PGE2 secretion, and mRNA transcription of genes involved in PG synthesis pathway in equine endometrial cells in vitro. In the in vivo study, we aimed to compare short-term effect of IUD inserted on day 0 (oestrus) with day 5–6 post-ovulation (the specific time when embryo reaches uterus after fertilization) on PG secretion from equine endometrium. To determine the long-term effect on PG synthase mRNA transcription, a single endometrial biopsy was taken only once within each group of mares at certain time points of the estrous cycle from mares placement with IUD on days 0 or 5–6 post-ovualtion.ResultsWe showed for the first time that the incubation of the endometrial cells with the presence of IUD altered the pattern of PG synthase mRNA transcription in equine epithelial and stromal endometrial cells. In vivo, in mares placement with IUD on day 0, PGE2 concentrations in blood plasma were upregulated between 1 and 6, and at 10 h after the IUD insertion, compared with the control mares (P < 0.05). Moreover, the decrease of PTGFS mRNA transcription on day 16- 18, associated with an elevation in PTGES mRNA transcription on day 20 -21 of the estrous cycle in endometrial biopsies collected from mares placement with IUD on days 5–6 suggest an antiluteolytic action of IUD during the estrous cycle.ConclusionWe conclude that the application of IUD may mimic the equine conceptus presence through the physical contact with the endometrium altering PG synthase transcription, and act as a potent modulator of endometrial PG secretion both in vitro and in vivo.

Prostaglandin F2α Affects the Cycle of Clock Gene Expression and Mouse Behavior.

Prostaglandins are bioactive compounds, and the activation of their receptors affects the expression of clock genes. However, the prostaglandin F receptor (Ptgfr) has no known relationship with biological rhythms. Here, we first measured the locomotor period lengths of Ptgfr-KO (B6.129-Ptgfrtm1Sna) mice and found that they were longer under constant dark conditions (DD) than those of wild-type (C57BL/6J) mice. We then investigated the clock gene patterns within the suprachiasmatic nucleus in Ptgfr-KO mice under DD and observed a decrease in the expression of the clock gene cryptochrome 1 (Cry1), which is related to the circadian cycle. Moreover, the expression of Cry1, Cry2, and Period2 (Per2) mRNA were significantly altered in the mouse liver in Ptgfr-KO mice under DD. In the wild-type mouse, the plasma prostaglandin F2α (PGF2α) levels showed a circadian rhythm under a 12 h cycle of light-dark conditions. In addition, in vitro experiments showed that the addition of PTGFR agonists altered the amplitude of Per2::luc activity, and this alteration differed with the timing of the agonist addition. These results lead us to hypothesize that the plasma rhythm of PGF2α is important for driving clock genes, thus suggesting the involvement of PGF2α- and Ptgfr-targeting drugs in the biological clock cycle.

In vitro maturation of African catfish (Clarias gariepinus, Burchell, 1822) oocytes results in viable larvae

In this study, we describe a protocol for in vitro maturation and ovulation of African catfish (Clarias gariepinus) postvitellogenic ovarian follicles, resulting in fertilizable and developmentally competent eggs. Such culture systems enable detailed studies of fish oocyte development and present a useful tool in assisted reproduction of fish. The maturation-inducing effects of 17α,20β-dihydroxy-4-pregnen-3-one (DHP), human chorionic gonadotropin (hCG) and recombinant human insulin-like growth factor 1 (rhIGF-1) were evaluated based on their ability to promote ooplasm translucency and germinal vesicle breakdown (GVBD), which indicates meiosis resumption. Among the tested groups, only follicles exposed to DHP underwent GVBD in a time-dependent manner, with the highest rates (>80%) observed within 10 to 12 h of incubation. Furthermore, adding rhIGF-1 or hCG prior to or during incubations with DHP did not influence the GVBD outcome. This suggested that the oocytes acquired maturational competence prior to isolation, as high concentrations of DHP (1 μg/ml) were alone sufficient to induce complete nuclear and cytoplasmic maturation; however, only 4 ± 3% of these oocytes ovulated in culture. To promote ovulation, we incorporated prostaglandins (PGs) as an additional incubation step following DHP-induced maturation. Exposure to both prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) led to increased rupture of the follicular layer, most notably in groups with 5 μg/ml PGF2α (71 ± 8%). In addition to identifying optimal hormonal stimulation, effects of culture media pH in combination with media supplementation—in the form of bovine serum albumin (BSA) and fetal bovine serum (FBS)—were considered. Compared to results in media with pH of 7.5, significantly less follicles matured and ovulated in a more alkaline environment (pH 8.5), regardless of supplement type and concentration. Although the presence of FBS (5–20%) slightly improved ovulation rates, this benefit was not reflected in the fertilization outcomes. In vitro matured and ovulated oocytes obtained in non-supplemented media (pH 7.5) maintained their developmental competence and were successfully fertilized. The hatching rate was 39%, after which the survival rate of larvae was 8% at 72 h post fertilization (hpf).

Efficacy of BAY 60-2770, a Soluble Guanylate Cyclase Activator, for Coronary Spasm in Animal Models.

Ischemia with non-obstructive coronary arteries (INOCA), caused by coronary artery spasm, has gained increasing attention owing to the poor quality of life of impacted patients. Therapeutic options to address INOCA remain limited, and developing new therapeutic agents is desirable. Here, we examined whether soluble guanylate cyclase (sGC) activators could be beneficial in preventing coronary spasms. In organ chamber experiments with isolated canine coronary arteries, prostaglandin F2 α -induced, endothelin-1-induced, 5-hydroxytryptamine-induced, and potassium chloride-induced contractions were suppressed by the sGC activator BAY 60-2770 (0.1, 1, and 10 nM). In isolated pig coronary arteries, BAY 60-2770 (0.1, 1, and 10 nM) could prolong the cycle length of phasic contractions induced by 3,4-diaminopyridine, as well as lower the peak and bottom tension of the contraction in a concentration-dependent manner. Additionally, BAY 60-2770 (1 pM-0.1 µM) evoked a concentration-related relaxation to a greater extent in small (first diagonal branch) coronary arteries than in large (left anterior descending) coronary arteries. In vasopressin-induced angina model rats, pretreatment with BAY 60-2770 (3 µg/kg) suppressed electrocardiogram S-wave depression induced by arginine vasopressin without affecting changes in mean blood pressure and heart rate. These findings suggest that BAY 60-2770 could be valuable in preventing both large and small coronary spasms. Therefore, sGC activators could represent a novel and efficacious therapeutic option for INOCA. SIGNIFICANCE STATEMENT: The soluble guanylate cyclase (sGC) activator BAY 60-2770 exerted antispastic effects on the coronary arteries in animal vasospasm models as proof-of-concept studies. These data can help to support potential clinical development with sGC activators, suitable for human use in patients with vasospastic angina.