Effect of undernutrition on the uterine environment during maternal recognition of pregnancy in sheep

2009 ◽  
Vol 21 (7) ◽  
pp. 869 ◽  
Author(s):  
C. Sosa ◽  
J. A. Abecia ◽  
M. Carriquiry ◽  
M. I. Vázquez ◽  
A. Fernández-Foren ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Types ◽  
Nuclear Antigen ◽  
Igf I ◽  
Uterine Environment ◽  
Cox 2 ◽  
Uterine Secretion ◽  
Maintenance Requirements ◽  
Proliferating Cell

The effects of pregnancy and undernutrition on endometrial gene expression were investigated in ewes fed all or half their maintenance requirements and killed on Day 14 of pregnancy or of the oestrous cycle. The endometrial expression of progesterone, oestrogen, oxytocin and interferon receptors (PR, ERα, OXTR and IFNAR, respectively), cyclo-oxygenase (COX)-2, proliferating cell nuclear antigen (PCNA), insulin-like growth factors (IGF)-I and -II, and IGF-1 receptor (IGF-1R) was studied by immunohistochemistry or real-time reverse transcription–polymerase chain reaction. The luminal epithelium of cyclic control ewes was devoid of PR staining and had relatively high levels of ERα, OXTR, COX-2 and IFNAR2. The presence of a conceptus decreased the in vitro uterine secretion of prostaglandin (PG) F2α and the expression of IFNAR2 in most cell types, and increased the gene expression of IGF-I and IGF-II. Undernutrition tended to increase ERα protein and gene, but decreased in vitro uterine secretion of PGE2 and the gene expression of IFNAR2 in cyclic ewes. There was no effect of undernutrition on pregnancy rates or the number of conceptuses recovered. Consistent with this, undernutrition of pregnant ewes did not have any effect on uterine gene expression. Moreover, in cases where changes were observed in cyclic ewes, these changes were negated when a conceptus was present.

scholarly journals Anastrozole and celecoxib for endometriosis treatment, good to keep them apart?

Reproduction ◽  
2013 ◽  
Vol 145 (2) ◽  
pp. 119-126 ◽  
Author(s):  
Carla N Olivares ◽  
Mariela A Bilotas ◽  
Analía G Ricci ◽  
Rosa Inés Barañao ◽  
Gabriela F Meresman
Keyword(s):  
Cell Proliferation ◽  
Nuclear Antigen ◽  
Gynecological Disease ◽  
Cox 2 ◽  
Benign Gynecological Disease ◽  
Surgically Induced ◽  
Free Women ◽  
Proliferating Cell

Endometriosis is a benign gynecological disease. Cyclooxygenase-2 (COX-2) and aromatase proteins have been shown to be overexpressed in eutopic endometrium from women suffering from this disease compared to disease-free women. Furthermore, inhibition of these molecules individually was demonstrated to have antiproliferative and proapoptotic effects both in vitro and in vivo in several models. In this study, the effect of combining celecoxib, a selective COX-2 inhibitor, and anastrozole, an aromatase inhibitor, on the implantation and growth of endometriotic like lesions in a murine model of endometriosis was evaluated. Endometriosis was surgically induced in female BALB/c mice. After 28 days of treatment with celecoxib, anastrozole, or their combination, animals were killed and lesions were counted, measured, excised, and fixed. Immunohistochemistry for proliferating cell nuclear antigen and CD34 was performed for assessment of cell proliferation and vascularization. TUNEL technique was performed for apoptosis evaluation. Celecoxib was the only treatment to significantly reduce the number of lesions established per mouse, their size and vascularized area. In addition, cell proliferation was significantly diminished and apoptosis was significantly enhanced by both individual treatments. When the therapies were combined, they reversed their effects. These results confirm that celecoxib and anastrozole separately decrease endometriotic growth, but when combined they might have antagonizing effects.

scholarly journals Potential chemopreventive, anticancer and anti-inflammatory properties of a refined artocarpin-rich wood extract of Artocarpus heterophyllus Lam.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Isaac J. Morrison ◽  
Jianan Zhang ◽  
Jingwen Lin ◽  
JeAnn E. Murray ◽  
Roy Porter ◽  
...  
Keyword(s):  
Gene Expression ◽  
Inflammatory Responses ◽  
Nuclear Antigen ◽  
Artocarpus Heterophyllus ◽  
Dss Colitis ◽  
Treatment And Prevention ◽  
Human Cytochrome ◽  
Anti Inflammatory ◽  
Proliferating Cell

AbstractColorectal cancer (CRC) represents the third leading cause of death among cancer patients below the age of 50, necessitating improved treatment and prevention initiatives. A crude methanol extract from the wood pulp of Artocarpus heterophyllus was found to be the most bioactive among multiple others, and an enriched extract containing 84% (w/v) artocarpin (determined by HPLC–MS–DAD) was prepared. The enriched extract irreversibly inhibited the activity of human cytochrome P450 CYP2C9, an enzyme previously shown to be overexpressed in CRC models. In vitro evaluations on heterologously expressed microsomes, revealed irreversible inhibitory kinetics with an IC50 value of 0.46 µg/mL. Time- and concentration-dependent cytotoxicity was observed on human cancerous HCT116 cells with an IC50 value of 4.23 mg/L in 72 h. We then employed the azoxymethane (AOM)/dextran sodium sulfate (DSS) colitis-induced model in C57BL/6 mice, which revealed that the enriched extract suppressed tumor multiplicity, reduced the protein expression of proliferating cell nuclear antigen, and attenuated the gene expression of proinflammatory cytokines (Il-6 and Ifn-γ) and protumorigenic markers (Pcna, Axin2, Vegf, and Myc). The extract significantly (p = 0.03) attenuated (threefold) the gene expression of murine Cyp2c37, an enzyme homologous to the human CYP2C9 enzyme. These promising chemopreventive, cytotoxic, anticancer and anti-inflammatory responses, combined with an absence of toxicity, validate further evaluation of A. heterophyllus extract as a therapeutic agent.

scholarly journals Enhancing droplet-based single-nucleus RNA-seq resolution using the semi-supervised machine learning classifier DIEM

2019 ◽  
Author(s):  
Marcus Alvarez ◽  
Elior Rahmani ◽  
Brandon Jew ◽  
Kristina M. Garske ◽  
Zong Miao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Cell Types ◽  
Supervised Machine Learning ◽  
Data Sets ◽  
Rna Seq ◽  
Novel Approach ◽  
Single Nucleus ◽  
Downstream Analysis

AbstractSingle-nucleus RNA sequencing (snRNA-seq) measures gene expression in individual nuclei instead of cells, allowing for unbiased cell type characterization in solid tissues. Contrary to single-cell RNA seq (scRNA-seq), we observe that snRNA-seq is commonly subject to contamination by high amounts of extranuclear background RNA, which can lead to identification of spurious cell types in downstream clustering analyses if overlooked. We present a novel approach to remove debris-contaminated droplets in snRNA-seq experiments, called Debris Identification using Expectation Maximization (DIEM). Our likelihood-based approach models the gene expression distribution of debris and cell types, which are estimated using EM. We evaluated DIEM using three snRNA-seq data sets: 1) human differentiating preadipocytes in vitro, 2) fresh mouse brain tissue, and 3) human frozen adipose tissue (AT) from six individuals. All three data sets showed various degrees of extranuclear RNA contamination. We observed that existing methods fail to account for contaminated droplets and led to spurious cell types. When compared to filtering using these state of the art methods, DIEM better removed droplets containing high levels of extranuclear RNA and led to higher quality clusters. Although DIEM was designed for snRNA-seq data, we also successfully applied DIEM to single-cell data. To conclude, our novel method DIEM removes debris-contaminated droplets from single-cell-based data fast and effectively, leading to cleaner downstream analysis. Our code is freely available for use at https://github.com/marcalva/diem.

scholarly journals An evaluation of antioxidant effects on recovery from postischemic acute renal failure.

1994 ◽  
Vol 4 (8) ◽  
pp. 1588-1597
Author(s):  
R A Zager ◽  
S M Fuerstenberg ◽  
P H Baehr ◽  
D Myerson ◽  
B Torok-Storb
Keyword(s):  
Renal Failure ◽  
Acute Renal Failure ◽  
Dna Fragmentation ◽  
Proliferating Cell Nuclear Antigen ◽  
Antigen Expression ◽  
Nuclear Antigen ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tubular Necrosis ◽  
Proliferating Cell

Xanthine oxidase (XO) activity and hydroxyl radical (.OH) formation are widely proposed mediators of renal reperfusion injury, potentially altering the severity of, and recovery from, postischemic acute renal failure. The goal of this study was to ascertain whether combination XO inhibitor (oxypurinol) and .OH scavenger (Na benzoate) therapy, given at the time of renal ischemia, alters the extent of: (1) tubular necrosis and filtration failure; (2) DNA fragmentation/apoptosis (assessed in situ by terminal deoxynucleotidyl transferase reactivity); (3) early tubular regenerative responses (proliferating cell nuclear antigen expression; (3H)thymidine incorporation); and (4) the rate and/or degree of functional and morphologic repair. The effects of XO inhibition, .OH scavengers, and "catalytic" iron (FeSO4) on human proximal tubular cell proliferation in vitro were also assessed with a newly established cell line (HK-2). Male Sprague-Dawley rats were subjected to 35 min of bilateral renal arterial occlusion with or without oxypurinol/benzoate therapy. These agents did not alter the extent of tubular necrosis or filtration failure, proliferating cell nuclear antigen expression or thymidine incorporation, or the rate/extent of renal functional/morphologic repair. DNA fragmentation did not precede tubular necrosis, and it was unaffected by antioxidant therapy. By 5 days postischemia, both treatment groups demonstrated regenerating epithelial fronds that protruded into the lumina. These structures contained terminal deoxynucleotidyl transferase-reactive, but morphologically intact, cells, suggesting the presence of apoptosis. Oxypurinol and .OH scavengers (benzoate; dimethylthiourea) suppressed in vitro tubular cell proliferation; conversely, catalytic Fe had a growth-stimulatory effect. These results suggest that: (1) XO inhibition/.OH scavenger therapy has no discernible net effect on postischemic acute renal failure; (2) DNA fragmentation does not precede tubular necrosis, suggesting that it is not a primary mediator of ischemic cell death; and (3) antioxidants can be antiproliferative for human tubular cells, possibly mitigating their potential beneficial effects.

Involvement of proliferating cell nuclear antigen (cyclin) in DNA replication in living cells

1989 ◽  
Vol 9 (1) ◽  
pp. 57-66
Author(s):  
M Zuber ◽  
E M Tan ◽  
M Ryoji
Keyword(s):  
Dna Polymerase ◽  
Proliferating Cell Nuclear Antigen ◽  
Living Cells ◽  
Nuclear Antigen ◽  
Plasmid Replication ◽  
Dna Polymerase Delta ◽  
Dna Polymerase Alpha ◽  
Proliferating Cell

Proliferating cell nuclear antigen (PCNA) (also called cyclin) is known to stimulate the activity of DNA polymerase delta but not the other DNA polymerases in vitro. We injected a human autoimmune antibody against PCNA into unfertilized eggs of Xenopus laevis and examined the effects of this antibody on the replication of injected plasmid DNA as well as egg chromosomes. The anti-PCNA antibody inhibited plasmid replication by up to 67%, demonstrating that PCNA is involved in plasmid replication in living cells. This result further implies that DNA polymerase delta is necessary for plasmid replication in vivo. Anti-PCNA antibody alone did not block plasmid replication completely, but the residual replication was abolished by coinjection of a monoclonal antibody against DNA polymerase alpha. Anti-DNA polymerase alpha alone inhibited plasmid replication by 63%. Thus, DNA polymerase alpha is also required for plasmid replication in this system. In similar studies on the replication of egg chromosomes, the inhibition by anti-PCNA antibody was only 30%, while anti-DNA polymerase alpha antibody blocked 73% of replication. We concluded that the replication machineries of chromosomes and plasmid differ in their relative content of DNA polymerase delta. In addition, we obtained evidence through the use of phenylbutyl deoxyguanosine, an inhibitor of DNA polymerase alpha, that the structure of DNA polymerase alpha holoenzyme for chromosome replication is significantly different from that for plasmid replication.

scholarly journals A Novel Gold Calreticulin Nanocomposite Based on Chitosan for Wound Healing in a Diabetic Mice Model

Nanomaterials ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 75 ◽  
Author(s):  
Sara Paola Hernández Martínez ◽  
Teodoro Iván Rivera González ◽  
Moisés Armides Franco Molina ◽  
Juan José Bollain y Goytia ◽  
Juan José Martínez Sanmiguel ◽  
...  
Keyword(s):  
Wound Healing ◽  
Topical Administration ◽  
Nuclear Antigen ◽  
Histological Evaluation ◽  
Mice Model ◽  
Fibroblast Migration ◽  
And Migration ◽  
Proliferating Cell

The development of new nanomaterials to promote wound healing is rising, because of their topical administration and easy functionalization with molecules that can improve and accelerate the process of healing. A nanocomposite of gold nanoparticles (AuNPs) functionalized with calreticulin was synthetized and evaluated. The ability of the nanocomposite to promote proliferation and migration was determined in vitro, and in vivo wound healing was evaluated using a mice model of diabetes established with streptozotocin (STZ). In vitro, the nanocomposite not affect the cell viability and the expression of proliferating cell nuclear antigen (PCNA). Moreover, the nanocomposite promotes the clonogenicity of keratinocytes, endothelial cells, and fibroblasts, and accelerates fibroblast migration. In vivo, mice treated with the nanocomposite presented significantly faster wound healing. The histological evaluation showed re-epithelization and the formation of granular tissue, as well as an increase of collagen deposition. Therefore, these results confirm the utility of AuNPs–calreticulin nanocomposites as potential treatment for wound healing of diabetic ulcers.

Influence of somatotropin on lipid metabolism and IGF gene expression in porcine adipose tissue

1992 ◽  
Vol 263 (4) ◽  
pp. E637-E645 ◽  
Author(s):  
C. K. Wolverton ◽  
M. J. Azain ◽  
J. Y. Duffy ◽  
M. E. White ◽  
T. G. Ramsay
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
Dietary Fat ◽  
Subcutaneous Adipose Tissue ◽  
Proliferation And Differentiation ◽  
Igf I ◽  
Systemic Factors ◽  
Subcutaneous Adipose ◽  
Acid Esterification

The present study was designed to evaluate the effects of porcine somatotropin (pST) treatment (2 mg/day) and dietary fat (10%) separately and in combination on the metabolic activity of subcutaneous adipose tissue, serum adipogenic activity, and insulin-like growth factor (IGF) gene expression within adipose tissue from growing 5- to 6-mo-old barrows. This study attempted to determine how these factors might contribute to the reported changes in adiposity of treated swine. Biopsies of adipose tissue were collected after 28 days of treatment following anesthesia with thiopental sodium (15 mg/kg iv). Somatotropin inhibited in vitro glucose oxidation and lipogenesis in adipose tissue but did not affect fatty acid esterification. Adipogenic activity of serum was not altered by pST treatment. Subcutaneous adipose tissue contained mRNA for IGF-I and -II, and pST administration increased the abundance of IGF-I mRNA. Dietary fat had no effect on these variables. Thus somatotropin reduces glucose metabolism in porcine subcutaneous adipose tissue. Preadipocyte proliferation and differentiation are not affected by somatotropin through its actions on systemic factors. Dietary fat provides no additional benefit in combination with pST administration to affect accretion of adipose tissue in growing swine.

scholarly journals 24 Validation of swine enteroids as a model to study Lawsonia intracellularis pathogenesis

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 21-22
Author(s):  
Ramya Lekha Medida ◽  
Talita P Resende ◽  
Connie Gebhart ◽  
Milena Saqui-Salces
Keyword(s):  
Gene Expression ◽  
Intestinal Epithelium ◽  
Culture Media ◽  
Paneth Cell ◽  
Nuclear Antigen ◽  
Atmospheric Conditions ◽  
Lawsonia Intracellularis ◽  
Swine Industry ◽  
Fatty Acid Binding

Abstract Lawsonia intracellularis is an obligate intracellular bacterium that causes proliferative ileitis, an enteric disease that costs more than $100 million annually to the US swine industry. Information about L. intracellualris pathogenesis is scarce due to lack of suitable in vitro models to study the infection-induced proliferative changes. L. intracellularis infection of the swine ileum has been shown to decrease the number of mucin-producing goblet cells and increase cell proliferation along with amplification of transient amplifying cells demonstrated by the expression of SOX9. The objective of this study was to validate the use of swine enteroids (three dimensional structures derived from adult intestinal stem cells) as a model to study the intestinal epithelial changes caused by L. intracellularis infection. Swine enteroids plated on transwell plate inserts to cover the surface area were infected with 108 L. intracellularis organisms in culture media per well and incubated at 37⁰C with atmospheric conditions of 8.0% oxygen, 8.8% carbon dioxide, and 83.2% nitrogen. Infected enteroids were collected after 7 days and gene expression levels of mucin 2 (MUC2), enterocyte marker fatty acid binding protein (FABP), endocrine marker chromogranin A (CGA), intestinal epithelium marker villin 1 (VIL1), paneth cell marker lysozyme (LYZ), proliferating cell nuclear antigen (PCNA) and SOX9 were measured. The expression of FABP and LYZ in L. intracellularis infected swine enteroids decreased by 50% and 20% respectively and the expression of SOX9 increased by 50%, with no significant changes for the levels of VIL1 and CGA. The changes observed in the infected swine enteroids reflect the profile of gene expression observed in L. intracellularis infected ileal tissue. Therefore, swine enteroids are a suitable in vitro system to study the dynamics of cell differentiation and proliferation of the intestinal epithelium induced by L. intracellularis infection.

Identification of the functional domain of p21WAF1/CIP1 that protects cells from cisplatin cytotoxicity

2005 ◽  
Vol 289 (3) ◽  
pp. F514-F520 ◽  
Author(s):  
Fang Yu ◽  
Judit Megyesi ◽  
Robert L. Safirstein ◽  
Peter M. Price
Keyword(s):  
Dominant Negative ◽  
Nuclear Antigen ◽  
Functional Domain ◽  
Binding Domains ◽  
Cdk2 Activity ◽  
Localization Signal ◽  
Induced Apoptosis ◽  
Proliferating Cell

The p21 cyclin-dependent kinase (cdk) inhibitor protects cells from cisplatin cytotoxicity in vivo and in vitro. However, the mechanism of protection is not known. Separate p21 domains are known to interact with several different proteins having proapoptotic functions. To investigate the mechanism of protection by p21, we have constructed adenoviruses encoding the different domains of p21. We were able to localize the protective activity to a region of 54 amino acids containing the cyclin-cdk interacting moiety. Other protein binding domains of p21, including the NH2-terminal procaspase-3 interactive region and the COOH-terminal region containing the proliferating cell nuclear antigen binding domain and the nuclear localization signal, had little protective effect on cisplatin cytotoxicity. The dependence of cisplatin cytotoxicity on cdk2 activity was also demonstrated because 1) cisplatin caused a marked increase in cdk2 activity, which was prevented by the p21 expression adenovirus, and 2) a cdk2 dominant-negative adenovirus also protected cells from cisplatin-induced apoptosis. Thus the data suggest that the mechanism of p21 protection is by direct inhibition of cdk2 activity and that cisplatin-induced apoptosis is caused by a cdk2-dependent pathway.

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