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1.
Am J Respir Crit Care Med ; 202(8): 1146-1158, 2020 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-32551816

RESUMO

Rationale: Antenatal inflammation with placental dysfunction is strongly associated with high bronchopulmonary dysplasia (BPD) risk in preterm infants. Whether antenatal or postnatal HIF (hypoxia-inducible factor) augmentation can preserve lung structure and function and prevent pulmonary hypertension after intrauterine inflammation is controversial.Objectives: To determine whether antenatal or postnatal prolyl-hydroxylase inhibitor (PHi) therapy increases lung HIF expression, preserves lung growth and function, and prevents pulmonary hypertension in a rat model of chorioamnionitis-induced BPD caused by antenatal inflammation.Methods: Endotoxin (ETX) was administered to pregnant rats by intraamniotic injection at Embryonic Day 20, and pups were delivered by cesarean section at Embryonic Day 22. Selective PHi drugs, dimethyloxalylglycine or GSK360A, were administered into the amniotic space at Embryonic Day 20 or after birth by intraperitoneal injection for 2 weeks. Placentas and lung tissue were collected at birth for morphometric and Western blot measurements of HIF-1a, HIF-2a, VEGF (vascular endothelial growth factor), and eNOS (endothelial nitric oxide synthase) protein contents. At Day 14, lung function was assessed, and tissues were harvested to determine alveolarization by radial alveolar counts, pulmonary vessel density, and right ventricle hypertrophy (RVH).Measurements and Main Results: Antenatal PHi therapy preserves lung alveolar and vascular growth and lung function and prevents RVH after intrauterine ETX exposure. Antenatal administration of PHi markedly upregulates lung HIF-1a, HIF-2a, VEGF, and eNOS expression after ETX exposure.Conclusions: HIF augmentation improves lung structure and function, prevents RVH, and improves placental structure following antenatal ETX exposure. We speculate that antenatal or postnatal PHi therapy may provide novel strategies to prevent BPD due to antenatal inflammation.


Assuntos
Displasia Broncopulmonar/tratamento farmacológico , Fator 1 Induzível por Hipóxia/metabolismo , Pulmão/efeitos dos fármacos , Peptídeo PHI/farmacologia , Prenhez , Aminoácidos Dicarboxílicos/farmacologia , Animais , Animais Recém-Nascidos , Western Blotting , Displasia Broncopulmonar/metabolismo , Displasia Broncopulmonar/patologia , Modelos Animais de Doenças , Endotoxinas/efeitos adversos , Endotoxinas/farmacologia , Feminino , Fator 1 Induzível por Hipóxia/efeitos dos fármacos , Imuno-Histoquímica , Técnicas In Vitro , Injeções Intralesionais , Pulmão/embriologia , Gravidez , Cuidado Pré-Natal , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/crescimento & desenvolvimento , Circulação Pulmonar/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Valores de Referência , Testes de Função Respiratória , Técnicas de Cultura de Tecidos
2.
J Anim Sci ; 98(6)2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32452520

RESUMO

A novel, non-terminal surgical procedure to remove a single placentome from the pregnant ewe for gene expression and histological analyses was recently developed in our laboratory. This technique allows for evaluation of nutritional insults on placental development at more than one stage of gestation using a single animal. Early attempts to develop a similar technique in cattle were met with complications due to inaccessibility of the gravid uterine horn because of its location and mass. One alternative is to collect a placentome from the contralateral uterine horn; however, the question remains as to whether gene expression varies among placentomes based on location relative to the fetus. Pregnant heifers were maintained on forage during early gestation and later moved into pens with a Calan gate system (American Calan, Northwood, NH). On gestational day (GD) 158, five heifers were assigned to receive a hay-based diet formulated to meet 100% of NRC requirements, and five heifers were fed 70% of NRC requirements until necropsy on GD244. At necropsy, a single representative placentome was selected for analysis from the antimesometrial side: (1) of the gravid uterine horn central to the amnion, (2) over the allantois immediately adjacent to the amnion, (3) in the tip of the gravid uterine horn, and (4) in the tip of the contralateral uterine horn. Mean placentome weight was greater (P < 0.05) for locations central to the amnion and allantois compared to locations within the tips of the ipsilateral and contralateral horns, respectively. Gene expression for angiogenic factors (FGF2, ODC1, VEGFA, and FLT1), nutrient transporters (SLC7A1 and SLC2A1), and factors associated with hormone action (ESR1, IGF1, IGFBP3, CSH1, and PAG1) were unaffected (P > 0.05) by dietary treatment or location of the placentome. Results indicate that location of the placentome in relation to the fetus does not impact gene expression, enhancing the efficacy of nonterminal methodologies for sampling gene expression in placentomes.


Assuntos
Bovinos/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Placenta/metabolismo , Prenhez , Útero/fisiologia , Animais , Dieta/veterinária , Feminino , Nutrientes , Placentação , Gravidez
3.
Nat Commun ; 11(1): 2111, 2020 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-32355283

RESUMO

Preterm birth (PTB) is the leading cause of perinatal mortality and newborn complications. Bile acids are recognized as signaling molecules regulating a myriad of cellular and metabolic activities but have not been etiologically linked to PTB. In this study, a hospital-based cohort study with 36,755 pregnant women is conducted. We find that serum total bile acid levels directly correlate with the PTB rates regardless of the characteristics of the subjects and etiologies of liver disorders. Consistent with the findings from pregnant women, PTB is successfully reproduced in mice with liver injuries and dysregulated bile acids. More importantly, bile acids dose-dependently induce PTB with minimal hepatotoxicity. Furthermore, restoring bile acid homeostasis by farnesoid X receptor activation markedly reduces PTB and dramatically improves newborn survival rates. The findings thus establish an etiologic link between bile acids and PTB, and open an avenue for developing etiology-based therapies to prevent or delay PTB.


Assuntos
Ácidos e Sais Biliares/sangue , Nascimento Prematuro/epidemiologia , Adolescente , Adulto , Animais , Tetracloreto de Carbono , Ácido Cólico/metabolismo , Modelos Animais de Doenças , Feminino , Hospitais , Humanos , Recém-Nascido , Hepatopatias/epidemiologia , Masculino , Camundongos , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/epidemiologia , Gravidez , Resultado da Gravidez , Prenhez , Nascimento Prematuro/sangue , Estudos Prospectivos , Transdução de Sinais , Adulto Jovem
4.
Plast Reconstr Surg ; 146(3): 552-562, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32459729

RESUMO

BACKGROUND: Traumatic muscle loss often results in poor functional restoration. Skeletal muscle injuries cannot be repaired without substantial fibrosis and loss of muscle function. Given its regenerative properties, the authors evaluated outcomes of fetal tissue-derived decellularized matrix for skeletal muscle regeneration. The authors hypothesized that fetal matrix would lead to enhanced myogenesis and suppress inflammation and fibrosis. METHODS: Composite tissue composed of dermis, subcutaneous tissue, and panniculus carnosus was harvested from the trunk of New Zealand White rabbit fetuses on gestational day 24 and from Sprague-Dawley rats on gestational day 18 and neonatal day 3, and decellularized using a sodium dodecyl sulfate-based negative-pressure protocol. Six, 10-mm-diameter, full-thickness rat latissimus dorsi wounds were created for each treatment, matrix was implanted (excluding the defect groups), and the wounds were allowed to heal for 60 days. Analyses were performed to characterize myogenesis, neovascularization, inflammation, and fibrosis at harvest. RESULTS: Significant myocyte ingrowth was visualized in both allogeneic and xenogeneic fetal matrix groups compared to neonatal and defect groups based on myosin heavy chain immunofluorescence staining. Microvascular networks were appreciated within all implanted matrices. At day 60, expression of Ccn2, Col1a1, and Ptgs2 were decreased in fetal matrix groups compared to defect. Neonatal matrix-implanted wounds failed to show decreased expression of Col1a1 or Ptgs2, and demonstrated increased expression of Tnf, but also demonstrated a significant reduction in Ccn2 expression. CONCLUSIONS: Initial studies of fetal matrices demonstrate promise for muscle regeneration in a rat latissimus dorsi model. Further research is necessary to evaluate fetal matrix for future translational use and better understand its effects.


Assuntos
Matriz Extracelular/genética , Regulação da Expressão Gênica , Desenvolvimento Muscular/genética , Músculo Esquelético/lesões , Prenhez , Engenharia Tecidual/métodos , Tecidos Suporte , Animais , Animais Recém-Nascidos , Western Blotting , Matriz Extracelular/metabolismo , Feminino , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Gravidez , RNA/genética , Coelhos , Ratos , Ratos Sprague-Dawley
5.
PLoS One ; 15(4): e0230422, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32271764

RESUMO

The inability of beef cattle to maintain full term pregnancies has become an economic concern for the beef industry. Herd management and nutritional improvements have alleviated environmental impacts on embryonic and fetal loss, yet additional gains can be made through genomic selection. The objectives of this study were to identify loci and gene-sets in crossbred beef heifers associated with the number of services required to become pregnant (TBRD) and heifer conception rate at first service (HCR1). Heifers (n = 709) from a commercial beef operation underwent one round of artificial insemination, before exposure to bulls for natural service for 50 days. Pregnancy and time of conception was determined by ultrasound 35 days after the breeding season. Heifers were genotyped using the GeneSeek (Lincoln, NE) Bovine GGP50K BeadChip prior to genome-wide association analyses (GWAA) conducted using an EIGENSTRAT-like model to identify loci associated (P < 1 × 10-5) with TBRD and HCR1. One locus was associated (P = 8.97 × 10-6) with TBRD on BTA19 and included the positional candidate gene ASIC2, which is differentially expressed in the endometrium of fertility classified heifers, and the positional candidate gene, SPACA3. Gene-set enrichment analyses using SNP (GSEA-SNP) data, was performed and identified one gene-set, oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen as enriched (NES = 3.15) with TBRD and contained nine leading edge genes that contributed to the enrichment of the gene set. The enriched gene-set is involved in catalyzing oxidation-reduction reactions, which have been associated with oxidative stressors impacting pregnancy success. No loci were associated nor gene-sets enriched with HCR1. Identification of loci, positional candidate genes, gene-sets and leading edge genes enriched for fertility facilitate genomic selection that allows producers to select for reproductively superior cattle, reduce costs associated with infertility, and increase percent calf crop.


Assuntos
Bovinos/genética , Loci Gênicos , Hibridização Genética/genética , Taxa de Gravidez , Prenhez , Reprodução/genética , Animais , Cruzamento , Quimera/genética , Cruzamentos Genéticos , Feminino , Fertilidade/genética , Fertilização/genética , Estudos de Associação Genética/veterinária , Técnicas de Genotipagem , Masculino , Polimorfismo de Nucleotídeo Único , Gravidez , Prenhez/genética
6.
Anim Sci J ; 91(1): e13378, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32329195

RESUMO

Interferon-tau (IFNT) regulates maternal recognition during early pregnancy in ruminants. The liver can serve as a hematopoietic organ, and it has immune functions. This study hypothesized whether mRNA and proteins of interferon-stimulated genes (ISGs) induced by early pregnancy are upregulated in maternal liver. Therefore, we determined the expression of interferon-stimulated gene 15-kDa protein (ISG15), 2',5'-oligoadenylate synthetase 1 (OAS1), myxovirus resistance protein 1 (MX1), interferon-gamma-inducible protein 10 (IP-10), and signal transducer and activator of transcription 1 (STAT1) in maternal livers during early pregnancy in sheep. Ovine livers were sampled on day 16 of the estrous cycle, and days 13, 16, and 25 of pregnancy, and expression of ISGs was detected by quantitative real-time PCR, Western blot, and immunohistochemistry analysis. Our results showed that there were increases in expression of the mRNA and proteins of ISG15, OAS1, IP-10, STAT1, and MX1 during early pregnancy. STAT1 protein was limited to the hepatocytes, and endothelial cells of proper hepatic arteries and hepatic portal veins. In conclusion, the upregulation of ISG15, OAS1, IP-10, STAT1, and MX1 proteins may be implicated in maternal hepatic immune adjustment and other functions during early pregnancy in sheep.


Assuntos
2',5'-Oligoadenilato Sintetase/genética , 2',5'-Oligoadenilato Sintetase/metabolismo , Quimiocina CXCL10/genética , Quimiocina CXCL10/metabolismo , Citocinas/genética , Citocinas/metabolismo , Expressão Gênica , Fígado/metabolismo , Proteínas de Resistência a Myxovirus/genética , Proteínas de Resistência a Myxovirus/metabolismo , Prenhez/metabolismo , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Ovinos/genética , Ovinos/fisiologia , Ubiquitinas/genética , Ubiquitinas/metabolismo , Animais , Ciclo Estral/genética , Ciclo Estral/metabolismo , Feminino , Fígado/imunologia , Gravidez , Prenhez/imunologia , Ovinos/imunologia , Regulação para Cima
7.
Gene ; 743: 144614, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32222532

RESUMO

Buffalo reproduction struggles with a high incidence of early embryonic mortality. Effective treatment and prevention strategies for this condition are not available due to lack of understanding of molecular pathways in early pregnancy of this species. In the present study, we have attempted to understand these molecular pathways by characterizing the endometrial transcriptomic profiles of pregnant buffalos during early pregnancy. For the transcriptome profiling, buffalo endometrial tissues of 29-36 days of pregnancy and of nonpregnant luteal phase were collected from the local slaughterhouse. We confirmed the status of pregnancy based on the crown vertebral length of the foetus. Total RNA was isolated and sequencing was performed using the Illumina nextseq platform. The raw reads were filtered and mapped to the Bos taurus UMD 3.1 reference genome assembly. An average of 24,597 genes was investigated for differential expression between the two groups. Transcriptome data identified a total of 450 differentially expressed genes (using a cut off value of log2 fold changes >2 and <-2) in early pregnancy in comparison to the nonpregnant group (Padj < 0.05). Among these, 270 genes were significantly upregulated and 180 genes were downregulated. The most impacted pathways were related to secretion, transport, ionic homeostasis, mitosis and negative regulation of viral processes. In conclusion, our study characterized a unique set of DEGs, during the early pregnancy of buffalo, which potentially modulate the endometrial environment to establish and maintain a successful pregnancy.


Assuntos
Búfalos/fisiologia , Endométrio/metabolismo , Prenhez/genética , RNA/metabolismo , Transcriptoma/fisiologia , Animais , Regulação para Baixo , Feminino , Gravidez , RNA/genética , RNA/isolamento & purificação , RNA-Seq , Regulação para Cima
8.
Exp Eye Res ; 193: 107981, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32088240

RESUMO

Preeclampsia (PE) is a hypertensive complication of pregnancy. Its cause is still unknown and it could be a risk factor for future ophthalmic problems. Retinal vascular bed alterations have been described as a consequence of PE, suggesting a retinopathy. Factors related to angiogenesis and vascular permeability, such as vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) or components of the renin angiotensin aldosterone system (RAAS), prorrenin/renin receptor ((P)RR) and angiotensin II type I receptor (AT1R) have been located in the retina, participating in other retinopathies, but it is unknown if they could participate in PE. Our aim was to elucidate whether VEGF, PEDF, (P)RR and AT1R could be modified during PE and during hypertension induced in rats with a history of PE. We used female Wistar rats and subrrenal aortic coarctation to induce PE, and after delivery, we induced a second hit by Nω-nitro-L-arginine methyl ester (L-NAME) administration. We measured blood pressure, proteinuria and pups development. In both models, eye fundal exploration and immunoblot for VEGF, PEDF, (P)RR and AT1R were performed. We found that the development of hypertension occurred faster in previously PE rats than in normal animals. VEGF, PEDF, (P)RR and AT1R were increased in PE, but in L-NAME-induced hypertension only (P)RR and AT1R were altered. Eye fundal data indicated that PE induced a level I retinopathy, but L-NAME induced a faster and more severe retinopathy in previously PE animals compared to previously normal pregnancy rats. These results indicate that PE predisposes to development of a faster and more severe retinopathy after a second hit. They also suggest that VEGF and PEDF seem to participate only in PE retinopathy, but in both models, RAAS components seem to have a more critical participation.


Assuntos
Indutores da Angiogênese/metabolismo , Retinopatia Hipertensiva/metabolismo , Pré-Eclâmpsia/metabolismo , Prenhez , Sistema Renina-Angiotensina/fisiologia , Retina/patologia , Vasos Retinianos/patologia , Animais , Permeabilidade Capilar , Modelos Animais de Doenças , Feminino , Retinopatia Hipertensiva/etiologia , Retinopatia Hipertensiva/patologia , Pré-Eclâmpsia/patologia , Gravidez , Ratos , Ratos Wistar , Retina/metabolismo , Vasos Retinianos/metabolismo
9.
Trop Anim Health Prod ; 52(5): 2205-2212, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32020411

RESUMO

With the development of standardization and scaling in the dairy farming industry, timely and accurate pregnancy diagnosis is required to improve the benefits of breeding by shortening the calving interval. However, the current pregnancy diagnostic methods cannot meet the requirements of the industry. Here, we review changes in the physiological indexes and in the function and morphological status of the reproductive organs of dairy cows at the early stages of pregnancy. In addition, the corresponding pregnancy diagnostic methods based on certain indexes are well development, and the pregnancy diagnostic approaches based on remote sensing and automation technology have become inevitable trends in the industry. These applications will reveal physiological regularities in pregnancy and benefit the detailed management of dairy cows during early pregnancy.


Assuntos
Bovinos/fisiologia , Indústria de Laticínios/métodos , Prenhez/fisiologia , Animais , Feminino , Gravidez
10.
Immunology ; 160(1): 52-63, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32052861

RESUMO

As a pineal gland hormone, melatonin acts through its receptors to modulate the immune system. The immune system is composed of primary and secondary organs, and immune organs are adapted to the presence of the fetal alloantigen during pregnancy. However, it is unclear whether melatonin affects maternal immune organs during early pregnancy in sheep. In this study, the ovine thymus, lymph node, spleen and liver were sampled at day 16 of the oestrous cycle, and at days 13, 16 and 25 of pregnancy. The expression of melatonin receptor 1A (MT1), melatonin receptor 1B (MT2) and cluster of differentiation 4 (CD4) was detected by quantitative real-time polymerase chain reaction, Western blot and immunohistochemistry experiments. Our results showed that during early pregnancy there was an upregulation of MT1 mRNA and protein in the thymus, lymph node and liver, and there was a downregulation in the spleen. The expression of MT2 mRNA and protein was increased in the thymus but decreased in the spleen and liver, and there was no significant change in the lymph node during early pregnancy. CD4 protein was upregulated in the thymus, lymph node and liver, but there were no significant changes in the spleen during early pregnancy. In conclusion, early pregnancy induces tissue-specific expression of MT1, MT2 and CD4, which may be due to the different functions of the thymus, lymph node, spleen and liver. Further, melatonin is involved in immune regulation of the maternal thymus, lymph node, spleen and liver during early pregnancy in sheep.


Assuntos
Antígenos CD4/metabolismo , Histocompatibilidade Materno-Fetal , Melatonina/metabolismo , Prenhez/imunologia , Receptores de Melatonina/metabolismo , Ovinos/imunologia , Animais , Feminino , Perfilação da Expressão Gênica , Tolerância Imunológica , Fígado/imunologia , Fígado/metabolismo , Linfonodos/imunologia , Linfonodos/metabolismo , Gravidez , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/imunologia , Baço/imunologia , Baço/metabolismo , Timo/imunologia , Timo/metabolismo , Regulação para Cima/imunologia
11.
Sci Rep ; 10(1): 2089, 2020 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-32034259

RESUMO

Despite their importance in mammalian reproduction, substances in the oxytocin-prostaglandins pathways have not been investigated in the horse placenta during most of pregnancy and parturition. Therefore, we quantified placental content of oxytocin (OXT), oxytocin receptor (OXTR), and prostaglandin E2 and F2 alpha during days 90-240 of pregnancy (PREG), physiological parturition (PHYS), and parturition with fetal membrane retention (FMR) in heavy draft horses (PREG = 13, PHYS = 11, FMR = 10). We also quantified OXTR and prostaglandin endoperoxide synthase-2 (PTGS2) mRNA expression and determined the immunolocalization of OXT, OXTR, and PTGS2. For relative quantification of OXT and OXTR, we used western blotting with densitometry. To quantify the prostaglandins, we used enzyme immunoassays. For relative quantification of OXTR and PTGS2, we used RT-qPCR. For immunolocalization of OXT, OXTR, and PTGS2, we used immunohistochemistry. We found that OXT was present in cells of the allantochorion and endometrium in all groups. PTGS2 expression in the allantochorion was 14.7-fold lower in FMR than in PHYS (p = 0.007). These results suggest that OXT is synthesized in the horse placenta. As PTGS2 synthesis is induced by inflammation, they also suggest that FMR in heavy draft horses may be associated with dysregulation of inflammatory processes.


Assuntos
Membranas Extraembrionárias/metabolismo , Cavalos/fisiologia , Ocitocina/metabolismo , Parto/metabolismo , Placenta/metabolismo , Prenhez/metabolismo , Prostaglandinas/metabolismo , Animais , Membranas Extraembrionárias/fisiologia , Feminino , Cavalos/metabolismo , Redes e Vias Metabólicas , Ocitocina/fisiologia , Parto/fisiologia , Placenta/fisiologia , Gravidez , Prenhez/fisiologia , Prostaglandinas/fisiologia
12.
Development ; 147(4)2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32001436

RESUMO

Proper organ development depends on coordinated communication between multiple cell types. Retinoic acid (RA) is an autocrine and paracrine signaling molecule essential for the development of most organs, including the lung. Despite extensive work detailing effects of RA deficiency in early lung morphogenesis, little is known about how RA regulates late gestational lung maturation. Here, we investigate the role of the RA catabolizing protein Cyp26b1 in the lung. Cyp26b1 is highly enriched in lung endothelial cells (ECs) throughout development. We find that loss of Cyp26b1 leads to reduction of alveolar type 1 cells, failure of alveolar inflation and early postnatal lethality in mouse. Furthermore, we observe expansion of distal epithelial progenitors, but no appreciable changes in proximal airways, ECs or stromal populations. Exogenous administration of RA during late gestation partially mimics these defects; however, transcriptional analyses comparing Cyp26b1-/- with RA-treated lungs reveal overlapping, but distinct, responses. These data suggest that defects observed in Cyp26b1-/- lungs are caused by both RA-dependent and RA-independent mechanisms. This work reports crucial cellular crosstalk during lung development involving Cyp26b1-expressing endothelium and identifies a novel RA modulator in lung development.


Assuntos
Epitélio/embriologia , Pulmão/embriologia , Alvéolos Pulmonares/embriologia , Ácido Retinoico 4 Hidroxilase/genética , Ácido Retinoico 4 Hidroxilase/fisiologia , Animais , Sistemas CRISPR-Cas , Diferenciação Celular , Células Endoteliais/citologia , Células Epiteliais/citologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Rim/embriologia , Camundongos , Camundongos Endogâmicos C57BL , Organogênese/efeitos dos fármacos , Gravidez , Prenhez , Transdução de Sinais , Células-Tronco/citologia , Tretinoína/farmacologia
13.
Reprod Domest Anim ; 55(4): 454-459, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31957075

RESUMO

Leptin regulates body weight and several physiological processes including reproduction. We evaluated the circulating levels of leptin in pregnant and non-pregnant bitches as well as their correlation with body weight, food intake and number of foetuses. Nineteen healthy German shepherd bitches were used and divided in two groups (pregnant n = 12 and non-pregnant n = 7). Blood samples were collected every 15 days starting from ovulation (Day 0) throughout pregnancy (pregnant group, P) or throughout luteal phase (non-pregnant group, NP) In pregnant bitches, leptin concentrations increased from the day of ovulation (1.32 ± 0.06 ng/ml) up to day 45 (1.51 ± 0.06 ng/ml; p < .01) and returned to baseline values from day 60 post-ovulation. In non-pregnant bitches, leptin concentrations remained constant throughout the whole observation period (estimated marginal mean ± SE=1.33 ± 0.38 ng/ml). Pairwise comparisons showed significant differences between P and NP at day 45 post-ovulation (p < .05). Multivariable models indicated that, controlling for time and litter size, there was a positive relationship between leptin concentration and BW (p < .05) although Pearson coefficients showed that the correlation between BW and leptin was only significant in NP animals at day 45 (r = 0.76, p < .05). The multivariable approach also suggested that, holding BW and time constant, leptin concentrations tend to increase as the number of puppies increased (p = .06). Our study supports indirectly the contribution of the feto-placental unit to the circulating maternal leptin concentration.


Assuntos
Cães/fisiologia , Leptina/sangue , Tamanho da Ninhada de Vivíparos , Prenhez/fisiologia , Animais , Peso Corporal/fisiologia , Cães/sangue , Feminino , Fase Luteal/fisiologia , Ovulação/fisiologia , Placenta , Gravidez , Prenhez/sangue
14.
Arq. bras. med. vet. zootec. (Online) ; 72(1): 102-106, Jan.-Feb. 2020. ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1088941

RESUMO

Multiple pregnancies in humans account for only 3% of pregnancies, 97-98% of which are twin pregnancies and the morbimortality is higher in the monochorionic twins when compared to dichorionic ones. The canine species is naturally multiparous, but the diagnosis of monochorionic twin pregnancy is not common. The objective of this report was to describe the ultrasonographic diagnosis of monochorionic twin pregnancies in two bitches [Pug (case 1) and Shih tzu (case 2)]. It was possible to verify the presence of one gestational vesicle containing two fetuses in each female by observing two heads or two bodies within the same placental site. These fetuses presented adequate viability and normal organogenesis. Their development was similar to the other fetuses. In case 1 they were stillborn and smaller than the other five live-born fetuses. The twins in case 2 were born alive, but they also appeared smaller when compared to the littermates. The gestational risks associated with this condition in pregnant bitches are still unknown, however, there are reports of fetal death in monochorionic pregnancies in this species. Therefore, ultrasonographic exam during pregnancy allows an early monochorionic diagnosis and monitoring the fetal viability could bring health benefits to both the female and the littermates.(AU)


As gestações múltiplas em humanos correspondem a apenas 3% das gestações, sendo 97-98% dessas gestações gemelares. Sabe-se que a morbimortalidade fetal é maior em gêmeos monocoriônicos do que nos dicoriônicos. A espécie canina é naturalmente multípara, mas o diagnóstico gestacional de gêmeos monocoriônicos não é comum. O objetivo deste relato é descrever o diagnóstico ultrassonográfico de gêmeos monocoriônicos em duas cadelas, sendo uma da raça Pug (caso 1) e outra da raça Shih-Tzu (caso 2). Foi possível verificar a presença de uma única vesícula gestacional contendo dois fetos em cada cadela, por meio da visibilização de duas cabeças ou de dois corpos dentro de uma mesma placenta. Esses fetos apresentavam viabilidade e organogênese adequadas e o grau de desenvolvimento era similar aos demais fetos da ninhada. No caso 1, os gêmeos nasceram mortos e de tamanho menor que os outros cinco fetos nascidos vivos. Os gêmeos do caso 2 nasceram vivos, mas também eram pequenos em relação aos irmãos da ninhada. Os riscos gestacionais associados a essa condição em cadelas ainda não são conhecidos, no entanto já existem relatos de morte fetal em gestação monocoriônica nessa espécie. Portanto, o exame ultrassonográfico durante a gestação permite um diagnóstico precoce da monocorionicidade, e a monitorização da viabilidade fetal pode trazer benefícios para a saúde da matriz, assim como para o restante da ninhada.(AU)


Assuntos
Animais , Feminino , Cães , Gravidez Múltipla , Prenhez , Ultrassonografia Pré-Natal/veterinária
15.
Animal ; 14(6): 1234-1240, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31907084

RESUMO

Progesterone (P4) plays a key role in pregnancy establishment and maintenance; during early pregnancy, P4 stimulates the production and release of uterine secretions necessary for conceptus growth prior to implantation; therefore, exogenous P4 supplementation may improve embryo development. This study evaluated the effects of supplementation during early pregnancy with long-acting injectable progesterone or altrenogest on embryonic characteristics of sows and gilts. Thus, a total of 32 sows and 16 gilts were used. On day 6 of pregnancy sows and gilts were allocated to one of the following groups: non-supplemented; supplemented with 20 mg of altrenogest, orally, from days 6 to 12 of pregnancy; supplemented with 2.15 mg/kg of long-acting injectable progesterone on day 6 of pregnancy. Animals were killed on day 28 of pregnancy, and ovulation rate, embryo survival, embryo weight, crown-to-rump length, uterine glandular epithelium and endometrial vascularization were assessed. Treatments had no effect on pregnancy rate, embryo survival or endometrial vascular density (P > 0.05). Non-supplemented gilts presented larger and heavier embryos compared to gilts from supplemented groups (P < 0.05). Sows in the altrenogest group presented larger and heavier embryos compared to non-supplemented sows and sows supplemented with long-acting injectable progesterone. In conclusion, supplementation of sows and gilts with progestagen from day 6 of pregnancy can be used as a means to improve embryo survival without deleterious effects.


Assuntos
Implantação do Embrião/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Prenhez , Suínos/fisiologia , Acetato de Trembolona/análogos & derivados , Animais , Suplementos Nutricionais , Embrião de Mamíferos , Endométrio , Feminino , Ovulação/fisiologia , Gravidez , Taxa de Gravidez , Prenhez/efeitos dos fármacos , Progestinas/administração & dosagem , Progestinas/farmacologia , Acetato de Trembolona/administração & dosagem , Acetato de Trembolona/farmacologia
16.
Development ; 147(4)2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-31988184

RESUMO

Integrin dimers α3/ß1, α6/ß1 and α6/ß4 are the mammary epithelial cell receptors for laminins, which are major components of the mammary basement membrane. The roles of specific basement membrane components and their integrin receptors in the regulation of functional gland development have not been analyzed in detail. To investigate the functions of laminin-binding integrins, we obtained mutant mice with mammary luminal cell-specific deficiencies of the α3 and α6 integrin chains generated using the Cre-Lox approach. During pregnancy, mutant mice displayed decreased luminal progenitor activity and retarded lobulo-alveolar development. Mammary glands appeared functional at the onset of lactation in mutant mice; however, myoepithelial cell morphology was markedly altered, suggesting cellular compensation mechanisms involving cytoskeleton reorganization. Notably, lactation was not sustained in mutant females, and the glands underwent precocious involution. Inactivation of the p53 gene rescued the growth defects but did not restore lactogenesis in mutant mice. These results suggest that the p53 pathway is involved in the control of mammary cell proliferation and survival downstream of laminin-binding integrins, and underline an essential role of cell interactions with laminin for lactogenic differentiation.


Assuntos
Integrinas/fisiologia , Lactação , Glândulas Mamárias Animais/fisiologia , Animais , Diferenciação Celular , Linhagem da Célula , Proliferação de Células , Sobrevivência Celular , Citoesqueleto/fisiologia , Progressão da Doença , Feminino , Deleção de Genes , Hormônios/fisiologia , Integrina alfa3/fisiologia , Integrina alfa6/fisiologia , Integrina beta1/fisiologia , Integrina beta4/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Knockout , Camundongos Mutantes , Mutação , Células-Tronco Neoplásicas/citologia , Análise de Sequência com Séries de Oligonucleotídeos , Ovário/fisiologia , Fenótipo , Gravidez , Prenhez , Prognóstico , Ligação Proteica , Multimerização Proteica
17.
Vet Radiol Ultrasound ; 61(2): 215-226, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31899939

RESUMO

Reproductive success is vital in sustaining free-ranging and managed bottlenose dolphin (Tursiops truncatus) populations. Ultrasonography is an invaluable, non-invasive tool in assessing the fetomaternal unit in humans and animals, including dolphins and horses. The purpose of this prospective longitudinal cohort study was to develop a protocol for fetomaternal ultrasonographic monitoring in dolphins and to report normal measurements and descriptive findings correlated with a positive outcome. From 2010 to 2017, serial ultrasonographic evaluations of 12 healthy dolphins were performed over the course of 16 pregnancies. A total of 203 ultrasound examinations were included in the study. Several metrics were accurate in predicting fetal age. Fetal biparietal diameter (BPD), thoracic width in dorsal and transverse planes, thoracic height in a sagittal plane, aortic diameter, and blubber thickness all demonstrated high correlation with gestational age (r > 0.94, P < .00001). Regional uteroplacental thickness significantly increased with each trimester (range 0.22-0.40 cm; P < .00011 cranial uterus, P < .00057 mid, and P < .000011 caudal). Lung:liver mean pixel intensity was 2.57 ± 0.46 (95% confidence interval 2.47-2.67). Ultrasonographic characteristics of normal pregnancy in dolphins are described and an equation for prediction of parturition date in Tursiops is reported: days to parturition = 348.16 - (26.03 × BPD(cm)) (R2  = 0.99). Future applications of these normal data will help identify in utero abnormalities indicative of fetal morbidity, and improve understanding of reproductive failure in wild and managed populations.


Assuntos
Golfinho Nariz-de-Garrafa , Prenhez , Ultrassonografia Pré-Natal/veterinária , Animais , Estudos de Coortes , Feminino , Desenvolvimento Fetal , Estudos Longitudinais , Gravidez , Estudos Prospectivos , Útero
18.
Exp Eye Res ; 192: 107915, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31911164

RESUMO

Limbal stem cells (LSCs), a subpopulation of limbal epithelial basal cells, are crucial to the homeostasis and wound healing of corneal epithelium. The identification and isolation of LSCs remains a challenge due to lack of specific LSCs biomarkers. In this study, Haematoxylin-eosin (HE), 4', 6-diamidino-2-phenylindole (DAPI), and immunohistochemistry (IHC) stains were performed on the pre- and post-natal limbus tissues of mice which has the advantage of more controllable in term of sampling age relative to human origin. By morphological analysis, we supported that there is an absence of the Palisades of Vogt (POV) in the mouse. The development of prenatal and neonatal cornea was dominated by its stroma, whereas after eyelids opened at P14, the corneal epithelial cells (CECs) quickly go stratification in response to the liquid-air interface. Based on IHC staining, we found that the expression of LSCs putative biomarkers in limbal epithelial basal cells appeared in chronological order as follows: Vim = p63 > CK14 > CK15 (where = represents same time; > represents earlier), and in corneal epithelial basal cells were weakened in chronological order as follows: Vim > p63 > CK15 > CK14, which might also represent the stemness degree. Furthermore, the dynamic spatial expression of the examined LSCs putative biomarkers during mouse development also implied a temporal restriction. The expression of Vim in epithelial cells of mouse ocular surface occurred during E12-E19 only. The expression of CK15 was completely undetectable in CECs after P14, whereas the others putative molecular markers of LSCs, such as p63 and CK14, still remained weak expression, suggesting that CK15 was suitable to serve as the mouse LSCs biomarkers after P14. In this study, our data demonstrated the dynamic spatiotemporal expression pattern of LSCs putative biomarkers in mouse was age-related and revealed the time spectrum of the expression of LSCs in mouse, which adds in our knowledge by understanding the dynamic expression pattern of biomarkers of stem cells relate to maintenance of their stemness.


Assuntos
Biomarcadores/metabolismo , Epitélio Anterior/metabolismo , Proteínas do Olho/metabolismo , Limbo da Córnea/embriologia , Limbo da Córnea/metabolismo , Prenhez , Células-Tronco/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Imuno-Histoquímica , Queratina-14/metabolismo , Queratina-15/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Gravidez , Análise Espaço-Temporal , Fatores de Tempo , Transativadores/metabolismo , Vimentina/metabolismo
19.
Ann Thorac Surg ; 109(3): 810-819, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31541634

RESUMO

BACKGROUND: Congenital heart disease (CHD) is associated with abnormal fetal brain development, a phenomenon that may be related to decreased cerebral oxygen delivery in utero. We used an artificial womb model to test the hypothesis that decreasing fetal oxygen delivery would reproduce physiologic changes identified in fetuses with CHD. METHODS: Experimental (hypoxemic) fetal lambs (mean gestational age, 111 ± 3 days; n = 4) and control animals (112 days; n = 5) were maintained in the artificial womb for a mean of 22 ± 6 days. Oxygen delivery was reduced to 15.6 ± 1.0 mL/kg/min in the hypoxemia animals versus 21.6 ± 2.0 mL/kg/min in the control animals. Blood chemistry analysis and sonographic evaluation were performed daily. An additional control group (n = 7) was maintained in utero and harvested for analysis at gestational age 134 ± 4 days. RESULTS: Physiologic variables were monitored continuously, and no statistical differences between the groups were identified. Fetal oxygen delivery and arterial partial pressure of oxygen were remarkably lower in the experimental group longitudinally. Increased umbilical artery and decreased middle cerebral artery resistance resulted in a lower cerebral to umbilical resistance ratio, similar to the brain sparing effect observed in human fetuses with CHD. Experimental brains were smaller than control brains in relation to the calvarium on magnetic resonance. CONCLUSIONS: Sustained hypoxemia in fetal sheep leads to altered cerebrovascular resistances and loss of brain mass, similar to human fetuses with CHD. This unique model provides opportunities to investigate the pathologic process underlying CHD-associated brain dysmaturity and to evaluate potential fetal neuroprotective therapies.


Assuntos
Encéfalo/patologia , Oxigenação por Membrana Extracorpórea/métodos , Hipóxia Fetal/terapia , Cardiopatias Congênitas/complicações , Oxigênio/sangue , Prenhez , Animais , Encéfalo/embriologia , Modelos Animais de Doenças , Feminino , Hipóxia Fetal/sangue , Hipóxia Fetal/etiologia , Idade Gestacional , Cardiopatias Congênitas/sangue , Cardiopatias Congênitas/diagnóstico , Imagem por Ressonância Magnética , Gravidez , Ovinos , Ultrassonografia Pré-Natal
20.
Theriogenology ; 143: 98-112, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31864010

RESUMO

The present study documented the expression and functional role of Fibroblast growth factors (FGFs) family and their receptors (Fibroblast growth factor receptor, FGFRs) in placenta (Cotyledon; COT, Caruncle; CAR) during different stages of pregnancy in water buffalo. Samples were collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2); Mid pregnancy (MP) and Late pregnancy (LP) while diestrus stage of oestrus cycle (NP) was taken as control. In addition, modulatory role of FGF2 on mRNA expression of von Willebrand factor (vWF), Proliferating cell nuclear antigen (PCNA), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage (CYP11A1), 3ß-hydroxysteroid dehydrogenase (3ßHSD) and BCL2 Associated X (BAX) were studied in cultured trophoblast cells (TCC), obtained from EP2. Real-time PCR (qPCR), Western blot, and immunohistochemistry were applied to investigate mRNA and protein expressions, and the localization of examined factors whereas, P4 secretion was assessed by RIA. The mRNA and protein expression of FGFs and its receptors were maximum (P < 0.05) during EP (EP1 and EP2) in COT. However, FGFR1 and FGFR4 were upregulated (P < 0.05) during EP2 and MP in COT. Similarly, the mRNA and protein expression of FGFs and its receptors were upregulated (P < 0.05) during all stages of pregnancy in CAR. FGF family members were localized in the cytoplasm of trophoblast cells as well as in fetal blood vessels. At 100 ng/ml dosage, FGF2 stimulated the transcript of vWF maximally (P < 0.05). P4 secretion in trophoblast cells treated with FGF2 was maximum with the highest dose at 72 h. These findings corroborate that FGF acts locally in the trophoblast cells to modulate steroid hormone viz. progesterone synthesis, promote angiogenesis and favors cell survivability indicating that this factor may play an essential role in the regulation of placental formation and function in buffalo.


Assuntos
Búfalos/fisiologia , Fatores de Crescimento de Fibroblastos/metabolismo , Regulação da Expressão Gênica/fisiologia , Placenta/metabolismo , Prenhez , Animais , Feminino , Fatores de Crescimento de Fibroblastos/genética , Neovascularização Fisiológica , Placenta/irrigação sanguínea , Gravidez , Prenhez/fisiologia
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