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Scientific Reports Mar 2024Biological samples are often frozen and stored for years and/or thawed multiple times, thus assessing their stability on long-term storage and repeated freeze-thaw...
Effect of very long-term storage and multiple freeze and thaw cycles on 11-dehydro-thromboxane-B and 8-iso-prostaglandin F levels in human urine samples by validated enzyme immunoassays.
Biological samples are often frozen and stored for years and/or thawed multiple times, thus assessing their stability on long-term storage and repeated freeze-thaw cycles is crucial. The study aims were to assess:-the long-term stability of two major enzymatic and non-enzymatic metabolites of arachidonic acid, i.e. urinary 11-dehydro-thromboxane-(Tx) B, 8-iso-prostaglandin (PG)F, and creatinine in frozen urine samples;-the effect of multiple freeze-thaw cycles. Seven-hundred and three urine samples measured in previously-published studies, stored at -40 °C, and measured for a second time for 11-dehydro-TxB (n = 677) and/or 8-iso-PGF (n = 114) and/or creatinine (n = 610) were stable over 10 years and the 2 measurements were highly correlated (all rho = 0.99, P < 0.0001). Urine samples underwent 10 sequential freeze-thaw cycles, with and without the antioxidant 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (10 mM); urinary 11-dehydro-TxB and creatinine were stable across all cycles (11-dehydro-TxB: 100.4 ± 21%; creatinine: 101 ± 7% of baseline at cycle ten; n = 17), while 8-iso-PGF significantly increased by cycle 6 (151 ± 22% of baseline at cycle ten, n = 17, P < 0.05) together with hydrogen peroxide only in the absence of antioxidant. Arachidonic acid metabolites and creatinine appear stable in human urines stored at -40 °C over 10 years. Multiple freeze-thaw cycles increase urinary 8-iso-PGF in urine samples without antioxidants. These data are relevant for studies using urine samples stored over long-term and/or undergoing multiple freezing-thawing.
Topics: Humans; Antioxidants; Arachidonic Acid; Creatinine; Freezing; Immunoenzyme Techniques; Prostaglandins F; Thromboxanes
PubMed: 38448541
DOI: 10.1038/s41598-024-55720-3 -
International Journal of Molecular... Feb 2024Prostaglandins are bioactive compounds, and the activation of their receptors affects the expression of clock genes. However, the prostaglandin F receptor () has no...
Prostaglandins are bioactive compounds, and the activation of their receptors affects the expression of clock genes. However, the prostaglandin F receptor () has no known relationship with biological rhythms. Here, we first measured the locomotor period lengths of (B6.129-) 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 mice under DD and observed a decrease in the expression of the clock gene cryptochrome 1 (), which is related to the circadian cycle. Moreover, the expression of , , and () mRNA were significantly altered in the mouse liver in mice under DD. In the wild-type mouse, the plasma prostaglandin F (PGF) 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 ::luc activity, and this alteration differed with the timing of the agonist addition. These results lead us to hypothesize that the plasma rhythm of PGF is important for driving clock genes, thus suggesting the involvement of PGF- and -targeting drugs in the biological clock cycle.
Topics: Mice; Animals; Dinoprost; Mice, Inbred C57BL; Circadian Rhythm; Biological Clocks; Suprachiasmatic Nucleus; Gene Expression; Cryptochromes
PubMed: 38339119
DOI: 10.3390/ijms25031841 -
BMC Veterinary Research Feb 2024Intrauterine 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...
BACKGROUND
Intrauterine 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) F and PGE 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.
RESULTS
We 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, PGE 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.
CONCLUSION
We 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.
Topics: Horses; Animals; Female; Dinoprostone; Prostaglandin-Endoperoxide Synthases; Prostaglandins F; Endometrium; Intrauterine Devices; RNA, Messenger
PubMed: 38310284
DOI: 10.1186/s12917-024-03889-0 -
The Journal of Reproduction and... Apr 2024Two studies were conducted to evaluate the effects of the follicular wave on ovarian function and fertility in dairy heifers and lactating cows. In study 1, the estrous...
Two studies were conducted to evaluate the effects of the follicular wave on ovarian function and fertility in dairy heifers and lactating cows. In study 1, the estrous cycle of the selected Holstein heifers was initially synchronized using two intra-muscular prostaglandin F (PGF) administrations 11 days apart. Heifers in group FFW (n = 14) received an intra-muscular 500 μg PGF administration on day 7 after detecting standing estrus, while Heifers in group SFW (n = 14) were administered PGF 13 days after detecting standing estrus. The pregnancy rates of FFW (n = 98) and SFW (n = 100) heifers were also determined 35-37 days after artificial insemination (AI). In Study 2, healthy Holstein lactating cows (n = 28) were randomly assigned to either the FFW (n = 14) or SFW (n = 14) groups. The estrous cycles of the cows were presynchronized using two intra-muscular administrations of PGF given 14 days apart. Then, the emergences of the follicular waves were induced using an Ovsynch protocol. The pregnancy rate of FFW (n = 99) versus SFW (n = 98) cows was also determined 35-37 days after AI. The ovulatory follicle and corpus luteum (CL) resulting from the ovulatory follicle of FFW were larger than those of the dominant follicle and the CL of SFW in dairy heifers and lactating cows. However, the pregnancy rate did not differ between the FFW and SFW groups in heifers and lactating cows 35-37 days after AI. In conclusion, although the characteristics of the ovulatory follicles in FFW versus SFW animals differed, the follicular wave in dairy heifers or lactating cows did not affect fertility.
Topics: Pregnancy; Cattle; Animals; Female; Progesterone; Lactation; Ovarian Follicle; Corpus Luteum; Fertility; Insemination, Artificial; Gonadotropin-Releasing Hormone; Estrus Synchronization; Dinoprost
PubMed: 38246626
DOI: 10.1262/jrd.2023-084 -
Oncology Research 2023Hepatocellular carcinoma (HCC) is a leading cause of death worldwide. Current therapies are effective for HCC patients with early disease, but many patients suffer...
Hepatocellular carcinoma (HCC) is a leading cause of death worldwide. Current therapies are effective for HCC patients with early disease, but many patients suffer recurrence after surgery and have a poor response to chemotherapy. Therefore, new therapeutic targets are needed. We analyzed gene expression profiles between HCC tissues and normal adjacent tissues from public databases and found that the expression of genes involved in lipid metabolism was significantly different. The analysis showed that AKR1C3 was upregulated in tumors, and high AKR1C3 expression was associated with a poorer prognosis in HCC patients. , assays demonstrated that the knockdown of AKR1C3 or the addition of the AKR1C3 inhibitor indomethacin suppressed the growth and colony formation of HCC cell lines. Knockdown of AKR1C3 in Huh7 cells reduced tumor growth . To explore the mechanism, we performed pathway enrichment analysis, and the results linked the expression of AKR1C3 with prostaglandin F2 alpha (PGF2α) downstream target genes. Suppression of AKR1C3 activity reduced the production of PGF2α, and supplementation with PGF2α restored the growth of indomethacin-treated Huh7 cells. Knockdown of the PGF receptor (PTGFR) and treatment with a PTGFR inhibitor significantly reduced HCC growth. We showed that indomethacin potentiated the sensitivity of Huh7 cells to sorafenib. In summary, our results indicate that AKR1C3 upregulation may promote HCC growth by promoting the production of PGF2α, and suppression of PTGFR limited HCC growth. Therefore, targeting the AKR1C3-PGF2α-PTGFR axis may be a new strategy for the treatment of HCC.
Topics: Humans; Aldo-Keto Reductases; Dinoprost; Carcinoma, Hepatocellular; Liver Neoplasms; Cell Line; Indomethacin; Aldo-Keto Reductase Family 1 Member C3
PubMed: 38188684
DOI: 10.32604/or.2023.030975 -
The Journal of Reproduction and... Feb 2024This study investigated the effects of timed artificial insemination (TAI) and equine chorionic gonadotropin (eCG) administration on lactating dairy cows under...
This study investigated the effects of timed artificial insemination (TAI) and equine chorionic gonadotropin (eCG) administration on lactating dairy cows under heat-stress conditions (average temperature-humidity index: 80). Timed artificial insemination was performed on the cows with (n = 57) or without (control, n = 41) supplementation with 500 IU of eCG at the day of PGF treatment using the CIDR-Ovsynch protocol. GnRH was administered, and a progesterone device (CIDR) was inserted on Day -10 of the treatment protocol. The CIDR was removed on Day -3, and the cows were treated with PGF. Two days later, a 2 GnRH injection was administered. Subsequently, AI was performed on Day 0 (16-20 h after the 2 GnRH injection), and pregnancy was diagnosed on Days 32 and 60. Plasma progesterone (P) concentrations were measured after AI. Results showed that the eCG group had a higher pregnancy per AI (P/AI) than the control group (43.9 vs. 12.2%, P = 0.002), which was also accompanied by elevated P levels. Four cows in the eCG group had multiple calves, representing 7.0 and 16.0% of the group and pregnant cows, respectively. In conclusion, 500 IU of eCG combined with CIDR-Ovsynch in lactating dairy cows under severe heat stress conditions successfully improved fertility. However, the protocol may have a slight risk of multiple births.
Topics: Pregnancy; Female; Cattle; Animals; Horses; Progesterone; Lactation; Dinoprost; Gonadotropin-Releasing Hormone; Estrus Synchronization; Insemination, Artificial; Chorionic Gonadotropin
PubMed: 38171736
DOI: 10.1262/jrd.2023-069 -
International Journal of Molecular... Dec 2023Peony pollen contains multiple nutrients and components and has been used as a traditional Chinese medicine with a long history, but the effect of the treatment of...
Peony pollen contains multiple nutrients and components and has been used as a traditional Chinese medicine with a long history, but the effect of the treatment of primary dysmenorrhea remains to be clarified. The aim of this study is to investigate the therapeutic effect of peony pollen on primary dysmenorrhea mice and the potential mechanism. A uterus contraction model in vitro and primary dysmenorrhea mice were used to evaluate the treatment effect of peony pollen on primary dysmenorrhea. The primary dysmenorrhea mice were treated with 62.5 mg/kg, 125 mg/kg, or 250 mg/kg of peony pollen, and the writhing response, latency period, histopathological changes in the uterus, prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, and infiltration of neutrophils and macrophages were investigated. Protein expression of interleukin 1 β (IL-1β), interleukin 6 (IL-6), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cyclooxygenase-2 (COX-2), microsomal prostaglandin-E synthase 1 (mPGEs-1), BCL2-Associated X (Bax), B-cell lymphoma-2 (BCL-2), caspase-3, and cleaved caspase-3 were detected by Western blot, and the oxidative stress related marker malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and reactive oxygen species (ROS) were evaluated. Peony pollen could attenuate spontaneous or oxytocin-induced uterus contractions in vitro. Moreover, peony pollen decreased the writhing times, prolonged the writhing latency, and reduced the pathological damage of uterine tissues. Furthermore, the inflammatory cell infiltration and the protein expression of IL-1β, IL-6, and NLRP3 were decreased. The COX-2/PGE2 pathway was inhibited; oxidative stress and apoptosis in the uterus also improved in the uterus of primary dysmenorrhea mice. Peony pollen exerts a positive effect on primary dysmenorrhea by inhibiting the inflammatory response and modulating oxidative stress and apoptosis by regulating the COX-2/PGE2 pathway.
Topics: Humans; Female; Mice; Animals; Dinoprostone; Dysmenorrhea; Cyclooxygenase 2; NLR Family, Pyrin Domain-Containing 3 Protein; Caspase 3; Paeonia; Interleukin-6; Dinoprost
PubMed: 38139073
DOI: 10.3390/ijms242417245 -
Scientific Reports Dec 2023The onset of productive life in dairy cattle, concomitant to parturition, is accompanied by a substantial decrease in fertility in comparison with non-lactating,...
The onset of productive life in dairy cattle, concomitant to parturition, is accompanied by a substantial decrease in fertility in comparison with non-lactating, nulliparous heifers. Follicular growth patterns differ between parous and nulliparous dairy cattle. Nulliparous heifers ovulate follicles with reduced antral age (RAA). This study aimed to exogenously reduce ovulatory follicle age in lactating dairy cows from 7 to 5 days old. Cows (n = 80) had their estrous cycles synchronized with the Double-Ovsynch program. At the final portion of this program, luteolysis was induced at either 5 (RAA) or 7 (Control) days following follicular wave emergence. RAA outcomes were estimated in comparison with Controls. RAA resulted in smaller follicles 2 days post-treatment. Despite lower serum concentrations of 17β-estradiol before treatment compared with Controls, the rate of increase in this hormone was greater for the RAA treatment. There was no difference in luteolysis rates between treatments. Proestrus (luteolysis onset to estrus onset) was prolonged in RAA cows. Cows with RAA had more intense estruses. Collectively, these results indicate that decreasing the age of the ovulatory follicle may improve the steroidogenic capacity of the dominant follicle and estrus expression intensity in lactating dairy cows.
Topics: Pregnancy; Cattle; Female; Animals; Luteolysis; Progesterone; Lactation; Ovulation; Estrus; Estradiol; Dinoprost; Gonadotropin-Releasing Hormone
PubMed: 38129551
DOI: 10.1038/s41598-023-50001-x -
Journal of Dairy Science May 2024Our objective was to determine the effect of a 200-µg dose of GnRH 25 d after previous artificial insemination (AI) in a Resynch-25 resynchronization program on... (Comparative Study)
Comparative Study
Our objective was to determine the effect of a 200-µg dose of GnRH 25 d after previous artificial insemination (AI) in a Resynch-25 resynchronization program on ovulatory response, circulating progesterone (P4) concentrations before and after treatment, and pregnancy per AI (P/AI) compared with a 100-µg dose in lactating Holstein cows. Experimental d 0 was considered the day of the previous AI. Lactating dairy cows (n = 3,240) with an average of 126 d in milk (DIM) and between 1 and 6 services were randomly assigned to receive 100 µg or 200 µg of GnRH on d 25 (GnRH25). On d 32 after AI, cows diagnosed nonpregnant with the presence of a corpus luteum (CL) detected by ultrasound (n = 1,249) received PGF treatments on d 32 and 33, followed by a GnRH 32 h later and AI 16 h after this last GnRH. Blood samples were collected on d 25, 32, and 34 to evaluate serum P4 concentrations. Transrectal ultrasonographic examination was performed on d 25 and 27 to assess ovulatory response to GnRH25. Cows were checked for pregnancy on d 32, 46, and 88 after AI. The larger dose of GnRH increased the overall proportion of cows that ovulated to the GnRH25 (25.0% for the 100-µg dose vs. 32.5% for the 200-µg dose). However, when cows were evaluated separately according to the pregnancy status on d 32 after AI, we found no treatment effect within cows pregnant and nonpregnant. Even though treatment increased the proportion of cows with serum P4 ≤0.42 ng/mL at the last GnRH treatment (G2; 86.2% for the 100-µg dose vs. 93.0% for the 200-µg dose), it did not affect P/AI on d 32, 46, and 88. Furthermore, a greater proportion of cows without a functional CL at GnRH25 had circulating P4 concentrations ≥1.00 ng/mL on d 32 and lower than 0.42 ng/mL on G2. These cows also had a greater P/AI on d 32, 46, and 88. In summary, the larger dose of GnRH on d 25 after AI did not increase the ovulatory response in nonpregnant cows and P/AI on d 32, 46, and 88 after AI after the Resynch-25 program. Additionally, nonpregnant cows without a functional CL at GnRH25 were better synchronized after the Resynch-25 protocol and had greater P/AI on d 32, 46, and 88 after timed-AI.
Topics: Animals; Cattle; Female; Pregnancy; Dinoprost; Estrus Synchronization; Fertility; Gonadotropin-Releasing Hormone; Insemination, Artificial; Lactation; Progesterone
PubMed: 38101731
DOI: 10.3168/jds.2023-23938 -
European Review For Medical and... Dec 2023Previous studies have shown that the disturbance of redox homeostasis plays a role in the pathogenesis of mood disorders. It is currently unclear whether oxidative...
OBJECTIVE
Previous studies have shown that the disturbance of redox homeostasis plays a role in the pathogenesis of mood disorders. It is currently unclear whether oxidative stress parameters can be used as biomarkers (state vs. trait). The aim of the present study was to investigate oxidative stress markers in patients with major depressive disorder (MDD) and bipolar disorder (BP) in acute depressive episodes and remission, and healthy individuals.
PATIENTS AND METHODS
Thirty-two patients with a diagnosis of MDD, 32 patients with a diagnosis of BP and 32 matched healthy controls were included in the study. We measured the serum levels of markers of oxidative damage, including 8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-Iso-prostaglandin F2α (8-iso-PGF2α; 8-isoprostane), and malondialdehyde (MDA), and also serum activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR) in both acute and remission phase, and in control group.
RESULTS
After controlling for the effects of age, sex, body mass index, and smoking status, serum 8-iso-PGF2α levels were significantly higher in both patient groups compared to controls, regardless of disease phase. The activities of GPX and GR were significantly lower in the acute phase in MDD patients compared to controls. Serum GR activity was lower in both acute and remission phase in MDD compared to BP.
CONCLUSIONS
Our results suggest that both MDD and BP are associated with a disturbed redox balance with a particularly pronounced increase in serum 8-iso-PGF2α levels in both groups and the presence of glutathione metabolism disorders in MDD patients. Further research is needed to confirm the importance of oxidative stress parameters as potential biomarkers of MDD and BP.
Topics: Humans; Bipolar Disorder; Dinoprost; Depressive Disorder, Major; Oxidative Stress; Biomarkers; Superoxide Dismutase; Malondialdehyde; 8-Hydroxy-2'-Deoxyguanosine; Glutathione Peroxidase
PubMed: 38095397
DOI: 10.26355/eurrev_202312_34588