Direct observation of hematopoietic progenitor chimerism in fetal freemartin cattle

<p>Abstract</p> <p>Background</p> <p>Cattle twins are well known as blood chimeras. However, chimerism in the actual hematopoietic progenitor compartment has not been directly investigated. Here, we analyzed fetal liver of chimeric freemartin cattle by combining a new anti-bovine CD34 antibody and Y-chromosome specific in situ hybridization.</p> <p>Results</p> <p>Bull-derived CD34⁺cells were detected in the liver of the female sibling (freemartin) at 60 days gestation. The level of bull-derived CD34⁺cells was lower in the freemartin than in its male siblings. Bull (Y⁺) and cow hematopoietic cells often occurred in separate clusters. Around clusters of Y⁺CD34⁺cells, Y⁺CD34^-cells were typically observed. The thymi were also strongly chimeric at 60 days of gestation.</p> <p>Conclusion</p> <p>The fetal freemartin liver contains clusters of bull-derived hematopoietic progenitors, suggesting clonal expansion and differentiation. Even the roots of the hematopoietic system in cattle twins are thus strongly chimeric from the early stages of fetal development. However, the hematopoietic seeding of fetal liver apparently started already before the onset of functional vascular anastomosis.</p

Prepartal high-energy feeding with grass silage-based diets does not disturb the hepatic adaptation of dairy cows during the periparturient period

The liver of dairy cow naturally undergoes metabolic adaptation during the periparturient period in response to the increasing demand for nutrients. The hepatic adaptation is affected by prepartal energy intake level and is potentially associated with inflammatory responses. lb study the changes in the liver function during the periparturient period, 16 cows (body condition score = 3.7 +/- 0.3, mean +/- standard deviation; parity = second through fourth) were allocated to a grass silage-based controlled-energy diet (104 MJ/d) or a high-energy diet (135 MJ/d) during the last 6 wk before the predicted parturition. Liver samples were collected by biopsy at 8 d before the predicted parturition (-8 d) and at 1 and 9 d after the actual parturition (1 and 9 d). The lipidomic profile of liver samples collected at -8 and 9 d was analyzed using ultra performance liquid chromatography-mass spectrometry-based lipidomics. Liver samples from all the time points were subjected to microarray analysis and the subsequent pathway analysis with Ingenuity Pathway Analysis software (Ingenuity Systems, Mountain View, CA). Prepartal energy intake level affected hepatic gene expression and lipidomic profiles prepartum, whereas little or no effect was observed postpartum. At. 8 d, hepatic lipogenesis was promoted by prepartal high-energy feeding through the activation of X receptor/retinoid X receptor pathway and through increased transcription of thyroid hormone-responsive (THRSP). Hepatic inflammatory and acute phase responses at -8 d were suppressed (z-score = -2.236) by prepartal high-energy feeding through the increase in the mRNA abundance of suppressor of cytokine signaling 3 (SOCS9) and the decrease in the mRNA abundance of interleukin 1 (IL1), nuclear factor kappa B 1 (NFKB1), apolipoprotein A1 (APOA1), serum amyloid A3 (SAA3), haptoglobin (HP), lipopolysaccharide-binding protein (LBP), and inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3). Moreover, prepartal high-energy feeding elevated hepatic concentrations of C18- (7%), C20- (17%), C21(26%), C23-sphingomyelins (26%), and total saturated sphingomyelin (21%). In addition, cows in both groups displayed increased lipogenesis at the gene expression level after parturition and alterations in the concentration of various sphingolipids between the first and last samplings. In conclusion, prepartal high-energy feeding promoted lipogenesis and suppressed inflammatory and acute phase responses in the liver before parturition, whereas only minor effects were observed after parturition.Peer reviewe

Expression of uterine oxytocin receptors and blood progesterone, 13,14-dihydro-15-keto-prostaglandin F2α, and ionized calcium levels in dystocic bitches

This study aimed to examine the etiology of canine dystocia by measuring the relative expression of oxytocin receptor (OXTR) mRNA and the concentration of serum progesterone, plasma PGF(2 alpha) metabolite (PGFM), and blood ionized calcium (iCa) near term and in dystocia. Altogether 58 bitches were included in this study, 41 of which underwent cesarean section (CS). The four CS groups were based on history: complete uterine inertia (CUI; n = 7), partial uterine inertia (PUI; n = 13), obstructive dystocia (OD; n = 10), and elective cesarean section (ECS; n = 11). An additional group of medically treated dystocia without CS (MD; n = 8) and a control group (C; n = 9) with normal parturition (without CS and medical treatment) were also formed. Blood samples were taken prior to CS or medical treatment. Progesterone concentrations were highest in the ECS and a significant difference (p 0.05) was observed in iCa concentrations between the groups. Relative OXTR mRNA expression was evaluated with real-time PCR from full-thickness uterine samples taken from the incision site during CS. The expression was highest in the ECS and the difference in expression was significant (p <0.05) between the ECS and the OD and between ECS and the combined dystocia (CUI, PUI, OD) groups (COMB2). The study supports previous reports of decreasing progesterone and increasing PGFM during prepartum luteolysis. Upregulation of OXTR occurs near term. In obstructive dystocia, a prolonged influence of oxytocin and uterine exhaustion may lead to downregulation of OXTR. Complete primary uterine inertia may have a different etiology as no clear decrease in OXTR was observed in CUI as in OD. It remains unclear if parturition ceases because of uterine inertia or if uterine inertia occurs because of ceased parturition and desensitization of receptors. (C) 2019 Elsevier Inc. All rights reserved.Peer reviewe

Prepartal overfeeding alters the lipidomic profiles in the liver and the adipose tissue of transition dairy cows

Introduction Physiological adaptations in the energy metabolism of dairy cows during the periparturient period are partly mediated by insulin resistance (IR), which may subsequently induce metabolic disorders postpartum. The molecular mechanisms underlying IR in dairy cows are largely unknown. Objective This study aimed to find a novel insight into the molecular mechanisms underlying IR in dairy cows during the periparturient period by analyzing the effects of prepartal overfeeding on the lipidomic profiles in the liver and adipose tissue (AT). Methods Sixteen cows were allocated to controlled-energy and high-energy feeding groups. Lipidomic profiling was conducted on liver and adipose tissue samples collected at 8 days prior to the predicted parturition, and 1 day (only AT) and 9 days after the actual parturition. Results Five ceramides (Cers) were identified to be significantly increased by prepartal overfeeding in AT in the analysis of the variance between groups within time points. Principal component-linear discriminant analysis showed that lipidomic profiles between the feeding groups were mainly characterized by phosphatidylcholines (PC), phosphatidylethanolamines (PE), lysophophosphatidylcholines (LysoPC), and lysophosphatidylethanolamines (LysoPE) in the liver, and by Cer, PE, and phosphatidylinositols (PI) in AT. Lipid class levels indicated that prepartal overfeeding elevated the concentration of PE, PI, LysoPC, LysoPE, and sphingomyelin in the liver, and increased the concentration of Cer in AT during the periparturient period. Conclusion Prepartal overfeeding significantly altered the concentrations of various sphingolipids, phospholipids, and lysophospholipids in the liver and AT of dairy cows during the periparturient period.Peer reviewe

Rasvalisä lypsylehmien herutusruokinnassa

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GSK3β Serine 389 Phosphorylation Modulates Cardiomyocyte Hypertrophy and Ischemic Injury

Prior studies show that glycogen synthase kinase 3β (GSK3β) contributes to cardiac ischemic injury and cardiac hypertrophy. GSK3β is constitutionally active and phosphorylation of GSK3β at serine 9 (S9) inactivates the kinase and promotes cellular growth. GSK3β is also phosphorylated at serine 389 (S389), but the significance of this phosphorylation in the heart is not known. We analyzed GSK3β S389 phosphorylation in diseased hearts and utilized overexpression of GSK3β carrying ser→ala mutations at S9 (S9A) and S389 (S389A) to study the biological function of constitutively active GSK3β in primary cardiomyocytes. We found that phosphorylation of GSK3β at S389 was increased in left ventricular samples from patients with dilated cardiomyopathy and ischemic cardiomyopathy, and in hearts of mice subjected to thoracic aortic constriction. Overexpression of either GSK3β S9A or S389A reduced the viability of cardiomyocytes subjected to hypoxia–reoxygenation. Overexpression of double GSK3β mutant (S9A/S389A) further reduced cardiomyocyte viability. Determination of protein synthesis showed that overexpression of GSK3β S389A or GSK3β S9A/S389A increased both basal and agonist-induced cardiomyocyte growth. Mechanistically, GSK3β S389A mutation was associated with activation of mTOR complex 1 signaling. In conclusion, our data suggest that phosphorylation of GSK3β at S389 enhances cardiomyocyte survival and protects from cardiomyocyte hypertrophy

GSK3β Serine 389 Phosphorylation Modulates Cardiomyocyte Hypertrophy and Ischemic Injury

Prior studies show that glycogen synthase kinase 3β (GSK3β) contributes to cardiac ischemic injury and cardiac hypertrophy. GSK3β is constitutionally active and phosphorylation of GSK3β at serine 9 (S9) inactivates the kinase and promotes cellular growth. GSK3β is also phosphorylated at serine 389 (S389), but the significance of this phosphorylation in the heart is not known. We analyzed GSK3β S389 phosphorylation in diseased hearts and utilized overexpression of GSK3β carrying ser→ala mutations at S9 (S9A) and S389 (S389A) to study the biological function of constitutively active GSK3β in primary cardiomyocytes. We found that phosphorylation of GSK3β at S389 was increased in left ventricular samples from patients with dilated cardiomyopathy and ischemic cardiomyopathy, and in hearts of mice subjected to thoracic aortic constriction. Overexpression of either GSK3β S9A or S389A reduced the viability of cardiomyocytes subjected to hypoxia–reoxygenation. Overexpression of double GSK3β mutant (S9A/S389A) further reduced cardiomyocyte viability. Determination of protein synthesis showed that overexpression of GSK3β S389A or GSK3β S9A/S389A increased both basal and agonist-induced cardiomyocyte growth. Mechanistically, GSK3β S389A mutation was associated with activation of mTOR complex 1 signaling. In conclusion, our data suggest that phosphorylation of GSK3β at S389 enhances cardiomyocyte survival and protects from cardiomyocyte hypertrophy