β2-Adrenoceptor desensitization in human alveolar macrophages induced by inhaled terbutaline in vivo is not counteracted by budesonide

2001 ◽  
Vol 100 (4) ◽  
pp. 451-457 ◽  
Author(s):  
Anita ZETTERLUND ◽  
Paul HJEMDAHL ◽  
Kjell LARSSON
Keyword(s):  
Alveolar Macrophages ◽  
Control Group ◽  
Transcriptional Level ◽  
Down Regulation ◽  
Bronchodilator Response ◽  
Treatment Groups ◽  
Before And After ◽  
Β2 Agonist

In vitro studies suggest that glucocorticoids may counteract β-agonist-induced desensitization of β-adrenoceptors by actions at the transcriptional level, but the clinical relevance of such findings is not clear. Oral terbutaline treatment decreases β-adrenoceptor sensitivity in alveolar macrophages in vivo. This effect is not counteracted by inhaled or orally taken steroids. We therefore examined whether inhaled terbutaline elicited a similar effect on β2-adrenoceptor sensitivity in alveolar macrophages, and if co-treatment with an inhaled steroid, budesonide, would prevent such down-regulation. Bronchoalveolar lavage (BAL) and lung function tests, including bronchodilator responses to inhaled terbutaline, were performed before and after 2 weeks of regular inhalation of terbutaline, 0.5 mg three times daily, and budesonide, 400 µg twice daily, or placebo, in 24 healthy volunteers. Four untreated subjects served as controls. A marked, approx. 90%, decrease in isoprenaline-induced cAMP accumulation in alveolar macrophages was found in both treatment groups after 2 weeks, with no difference between placebo and budesonide (P = 0.45). In the untreated control group, cAMP responses to both isoprenaline and prostaglandin E1 tended to be lower on the second occasion. A limited, non-specific desensitization of adenylate cyclase activity thus contributed to the marked desensitization elicited by terbutaline inhalations. The bronchodilator response to inhaled terbutaline did not change after treatment in any of the three groups (F = 0.9, P = 0.50). In conclusion, inhalation of a β-agonist induced marked down-regulation of β2-adrenoceptor sensitivity in alveolar macrophages in vivo without influencing the bronchodilator response to a β2-agonist in healthy subjects. Co-treatment with an inhaled steroid failed to counteract the desensitization of alveolar macrophage β2-adrenoceptors.

scholarly journals The Diverse Potential of Gluten from Different Durum Wheat Varieties in Triggering Celiac Disease: A Multilevel In Vitro, Ex Vivo and In Vivo Approach

Nutrients ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3566
Author(s):  
Federica Gaiani ◽  
Sara Graziano ◽  
Fatma Boukid ◽  
Barbara Prandi ◽  
Lorena Bottarelli ◽  
...  
Keyword(s):  
Immune Response ◽  
Celiac Disease ◽  
Durum Wheat ◽  
Ex Vivo ◽  
Diet Group ◽  
Control Group ◽  
Wheat Varieties ◽  
Before And After

The reasons behind the increasing prevalence of celiac disease (CD) worldwide are still not fully understood. This study adopted a multilevel approach (in vitro, ex vivo, in vivo) to assess the potential of gluten from different wheat varieties in triggering CD. Peptides triggering CD were identified and quantified in mixtures generated from simulated gastrointestinal digestion of wheat varieties (n = 82). Multivariate statistics enabled the discrimination of varieties generating low impact on CD (e.g., Saragolla) and high impact (e.g., Cappelli). Enrolled subjects (n = 46) were: 19 healthy subjects included in the control group; 27 celiac patients enrolled for the in vivo phase. Celiacs were divided into a gluten-free diet group (CD-GFD), and a GFD with Saragolla-based pasta group (CD-Sar). The diet was followed for 3 months. Data were compared between CD-Sar and CD-GFD before and after the experimental diet, demonstrating a limited ability of Saragolla to trigger immunity, although not comparable to a GFD. Ex vivo studies showed that Saragolla and Cappelli activated immune responses, although with great variability among patients. The diverse potential of durum wheat varieties in triggering CD immune response was demonstrated. Saragolla is not indicated for celiacs, yet it has a limited potential to trigger adverse immune response.

scholarly journals Synbiotics Containing Nanoprebiotics: A Novel Therapeutic Strategy to Restore Gut Dysbiosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Liang Hong ◽  
Sang-Mok Lee ◽  
Whee-Soo Kim ◽  
Yun-Jaie Choi ◽  
Seo-Ho Oh ◽  
...  
Keyword(s):  
Microbial Diversity ◽  
Control Group ◽  
Treatment Groups ◽  
Gut Dysbiosis ◽  
Intracellular Stimulation ◽  
Novel Strategy ◽  
Experimental Treatments ◽  
New Formulation

A new formulation, nanoprebiotics [e.g., phthalyl pullulan nanoparticles (PPNs)], was demonstrated to enhance the antimicrobial activity of probiotics [e.g., Lactobacillus plantarum (LP)] in vitro through intracellular stimulation better than that by backbone prebiotics, which are commonly used. In this study, we aimed to investigate whether this combination would exert distinct effects as synbiotics in vivo. Synbiotics combinations of LP, pullulan, and PPNs were used as experimental treatments in a dysbiosis-induced murine model, and their restorative effect was assessed using pathogen Escherichia coli K99 challenge. Our results showed that the E. coli infection was suppressed markedly in the experimental group fed with synbiotics containing PPNs. In addition, the decrease in serum endotoxin level after synbiotics treatment suggested the reinforcement of the gut barrier. Comparison of treatment groups, including a normal control group, showed that synbiotics containing PPNs increased microbial diversity, which is a representative parameter of healthy status. Furthermore, distinct from probiotics treatment alone, synbiotics showed additive effects of enrichment of several well-known beneficial bacteria such as Lactobacillus, Bifidobacterium, and other butyrate-producing bacteria including Faecalibacterium. Collectively, our results indicate that synbiotics containing PPNs are effective at restoring gut dysbiosis, suppressing pathogenic infection, and increasing microbial diversity, suggesting that synbiotics with nanoprebiotics have the potential to be a novel strategy for ameliorating gut dysbiosis and infectious diseases.

scholarly journals Comparative in vitro study of root roughness after instrumentation with ultrasonic and diamond tip sonic scaler

2006 ◽  
Vol 14 (2) ◽  
pp. 124-129 ◽  
Author(s):  
Fernanda Vieira Ribeiro ◽  
Renato Correa Viana Casarin ◽  
Francisco Humberto Nociti Júnior ◽  
Enilson Antônio Sallum ◽  
Antonio Wilson Sallum ◽  
...  
Keyword(s):  
Surface Roughness ◽  
In Vitro Study ◽  
Root Surface ◽  
Control Group ◽  
Human Teeth ◽  
Treatment Groups ◽  
Before And After ◽  
Root Surfaces ◽  
Scanning Electron

OBJECTIVE: The purpose of this study was to evaluate the root surface roughness after instrumentation with hand curette and diamond-coated sonic and universal ultrasonic tips. MATERIALS AND METHODS: Forty root surfaces of human teeth were randomly assigned to four treatment groups: control group (without instrumentation), curette instrumentation, ultrasonic instrumentation with universal tip and sonic instrumentation with diamond-coated tip. Each sample was instrumented with fifteen strokes. Before and after instrumentation, surface roughness was measured. In addition, the root surface topography was examined after treatment under the scanning electron microscope. RESULTS: Significant statistical differences (p <0.05) were observed when comparing the control group (0.48±0.07mm) to the treated groups (hand - 1.246±0.279mm, ultrasonic - 1.468±0.177mm and sonic instrumentation - 1.576±0.20mm). The highest roughness was produced by diamond-coated sonic tip and by ultrasonic universal tip (p >0.05). CONCLUSION: The diamond-coated tip with sonic scaler instrumentation and ultrasonic instrumentation produce similar root surface roughness, higher than curette instrumentation.

scholarly journals Lysophosphatidic acid suppresses endothelial cell CD36 expression and promotes angiogenesis via a PKD-1–dependent signaling pathway

Blood ◽  
2011 ◽  
Vol 117 (22) ◽  
pp. 6036-6045 ◽  
Author(s):  
Bin Ren ◽  
James Hale ◽  
Sowmya Srikanthan ◽  
Roy L. Silverstein
Keyword(s):  
Endothelial Cell ◽  
Signaling Pathway ◽  
Lysophosphatidic Acid ◽  
Malignant Tumors ◽  
Transcriptional Level ◽  
Dependent Manner ◽  
Down Regulation ◽  
Thrombospondin 1

Abstract In pathologic settings including retinal ischemia and malignant tumors, robust angiogenesis occurs despite the presence in the microenvironment of antiangiogenic proteins containing thrombospondin structural homology (TSR) domains. We hypothesized that antiangiogenesis mediated by TSR-containing proteins could be blunted by localized down-regulation of their cognate receptor on microvascular endothelial cells (MVECs), CD36. Through screening a panel of endothelial cell agonists, we found that lysophosphatidic acid (LPA) dramatically down-regulated CD36 surface expression on primary MVECs. LPA is a lipid-signaling mediator known to have proangiogenic activity, but the mechanisms are largely unknown. We observed that LPA caused CD36 down-regulation in a dose- and time-dependent manner and was long lasting. Down-regulation occurred at the transcriptional level via a signaling pathway involving specific LPA receptors and protein kinase D. LPA-induced MVEC CD36 repression significantly attenuated in vitro antiangiogenic responses to thrombospondin-1, including blockade of migration, tube formation, and VEGFR-2 signaling in response to fibroblast growth factor-2. In vivo relevance was demonstrated by showing that LPA abrogated thrombospondin-1–mediated inhibition of neovascularization of Matrigel plugs implanted in mice. Our data thus indicate that the proangiogenic mechanism of LPA may in part be via switching off the antiangiogenic switch mediated by TSR proteins and CD36.

Apelin-13 deteriorates hypertension in rats after damage of the vascular endothelium by ADMA

2013 ◽  
Vol 91 (9) ◽  
pp. 708-714 ◽  
Author(s):  
Xue Han ◽  
Dong-Liang Zhang ◽  
Dao-Xin Yin ◽  
Qi-Dong Zhang ◽  
Wen-Hu Liu
Keyword(s):  
Endothelial Dysfunction ◽  
Treated Group ◽  
Control Group ◽  
Transmission Electron ◽  
Before And After ◽  
Pass Through ◽  
Mlc Phosphorylation ◽  
And Control

Asymmetric dimethylarginine (ADMA) is a risk factor for endothelial dysfunction. The polypeptide apelin has biphasic effects on blood vessels in vivo and in vitro. We investigated the effect of apelin-13 on ADMA-damaged vessels. Rats were divided among ADMA-treated and control groups, which were treated with ADMA (10 mg·(kg body mass)−1·day−1) or saline, respectively, for 4 weeks. Systolic blood pressure (SBP) was measured before and after the injection of apelin-13. The ultrastructure of endothelial cells in caudal arteries was examined using transmission electron microscopy. The reactivities of isolated caudal artery rings were observed after exposure to apelin-13, and myosin light chain (MLC) phosphorylation was assessed by immunohistochemistry in rings treated with or without apelin-13. ADMA induced hypertension and endothelial dysfunction. After injection of apelin-13, SBP declined in the control group but was elevated in the ADMA-treated group. In vitro, apelin-13 caused relaxation in rings in the control group, but it contracted rings in the ADMA-treated group. Apelin-13 promoted MLC phosphorylation in vascular smooth muscle cells (VSMCs) in the ADMA group. These results indicate that apelin-13 might pass through ADMA-damaged endothelium and act on VSMCs to increase MLC phosphorylation, thus contributing to vasoconstriction and exacerbating hypertension.

scholarly journals Selenide Chitosan Sulfate Improved The Hepatocyte Activity, Growth Performance, and Anti-Oxidation Ability by Activating The Thioredoxin Reductase of Chickens In Vitro and In Vivo

2020 ◽  
Author(s):  
Lele Hou ◽  
Huiling Qiu ◽  
Lianqin Zhu ◽  
Yufeng Huang ◽  
Shansong Gao ◽  
...  
Keyword(s):  
Growth Performance ◽  
Culture Medium ◽  
Thioredoxin Reductase ◽  
Dietary Supplementation ◽  
Control Group ◽  
Treatment Groups ◽  
Thioredoxin Reductase 1 ◽  
Specific Pathogen

Abstract Background: There are very few studies on the synergy effects of biological antioxidant activity on selenium (Se) and sulfate. This study evaluated the effect of selenide chitosan sulfate (LS-COS-Se) on the hepatocytes activity, growth performance, and anti-oxidation ability by activating the thioredoxin reductase (TrxR) system of specific pathogen free (SPF) chickens in vitro and in vivo. Methods: The hepatocytes were obtained in vitro and a total of 240 SPF White Leghorns chickens (7 days of age and body weight of 45.0 ± 2.0 g) were collected in vivo. The hepatocytes and chickens were randomly allocated into six treatment groups: control group; chitosan (COS) group; sodium selenite (Na2SeO3) group; selenide chitosan (COS-Se) group; chitosan sulfate (LS-COS) group; LS-COS-Se group. After 24 h, the culture medium and hepatocytes were collected and preserved respectively for analyzing the metabolic activity of hepatocytes. Gowth performance was evaluated and chickens were euthanized to obtain plasma and liver tissue to measure antioxidant associated parameter on days 14 and 28. Results: The experiment in vitro showed that the activities of TrxR, superoxide dismutase (SOD), catalase (CAT) in culture medium and the levels of thioredoxin reductase 1 (TrxR-1) and thioredoxin reductase 3 (TrxR-3) mRNA in hepatocytes in LS-COS-Se group were significantly higher (P < 0.05), but the content of malondialdehyde (MDA) and the activity of lactate dehydrogenase (LDH) significantly decreased (P < 0.05) than those in control, COS and LS-COS groups. Compared with Na2SeO3 and COS-Se groups, the levels of TrxR-1 and TrxR-3 mRNA in hepatocytes and the activity of SOD in culture medium significantly increased in LS-COS-Se group (P < 0.05). The experiment in vivo showed that the baby weight on 14d and 28d, the activities of TrxR, SOD and anti-superoxide anion radical (AntiO2-) in plasma and the levels of TrxR-1 and TrxR-3 mRNA in liver of dietary supplementation with LS-COS-Se were significantly higher than those in control, COS and LS-COS groups (P < 0.05). The activities of TrxR and SOD in plasma of dietary supplementation with LS-COS-Se were significantly higher than those of Na2SeO3 group and COS-Se group (P < 0.05). Conclusion: LS-COS-Se as potential antioxidant improved the hepatocytes activity, growth performance, and anti-oxidation ability by activating the TrxR system of SPF chickens in vitro and in vivo. The better biological activity of LS-COS-Se was mainly due to the synergistic effect of Se and sulfate on TrxR system.

Morphogenesis-related gene-expression profile in porcine oocytes before and after in vitro maturation

Zygote ◽  
2017 ◽  
Vol 25 (3) ◽  
pp. 331-340 ◽  
Author(s):  
Joanna Budna ◽  
Adrian Chachuła ◽  
Dominika Kaźmierczak ◽  
Marta Rybska ◽  
Sylwia Ciesiółka ◽  
...  
Keyword(s):  
Control Group ◽  
Affymetrix Microarray ◽  
Brilliant Cresyl Blue ◽  
Cresyl Blue ◽  
Before And After ◽  
Porcine Oocyte ◽  
Developmental Capacity ◽  
A Cell

SummaryMammalian oocyte maturation is achieved when oocytes reach metaphase II (MII) stage, and accumulate mRNA and proteins in the cytoplasm following fertilization. It has been shown that oocytes investigated before and after in vitro maturation (IVM) differ significantly in transcriptomic and proteomic profiles. Additionally, folliculogenesis and oogenesis is accompanied by morphogenetic changes, which significantly influence further zygote formation and embryo growth. This study aimed to determine new transcriptomic markers of porcine oocyte morphogenesis that are associated with cell maturation competence. An Affymetrix microarray assay was performed on an RNA template isolated from porcine oocytes before (n = 150) and after (n = 150) IVM. The brilliant cresyl blue (BCB) staining test was used for identification of cells with the highest developmental capacity. DAVID (Database for Annotation, Visualization, and Integrated Discovery) software was used for the extraction of the genes belonging to a cell morphogenesis Gene Ontology group. The control group consisted of freshly isolated oocytes. In total, 12,000 different transcripts were analysed, from which 379 genes were downregulated and 40 were upregulated in oocytes following IVM. We found five genes, SOX9, MAP1B, DAB2, FN1, and CXCL12, that were significantly upregulated in oocytes after IVM (in vitro group) compared with oocytes analysed before IVM (in vivo group). In conclusion, we found new transcriptomic markers of oocyte morphogenesis, which may be also recognized as significant mediators of cellular maturation capacity in pigs. Genes SOX9, MAP1B, DAB2, FN1, and CXCL12 may be involved in the regulation of the MII stage oocyte formation and several other processes that are crucial for porcine reproductive competence.

scholarly journals Role of Vitamin D on the Inhibition of Gastrin Production After Cisplatin Treatment

2000 ◽  
Vol 7 (3) ◽  
pp. 115-119 ◽  
Author(s):  
Ying Wang ◽  
Surinder K. Aggarwal ◽  
Will Kopachik
Keyword(s):  
Vitamin D ◽  
Active Form ◽  
Treatment Groups ◽  
Calcium Supplements ◽  
Blot Technique ◽  
Before And After ◽  
Male Wistar Rats

In rats cisplatin induces hypocalcemia, bloating of the stomach, and ulceration ameliorated through calcium supplements. This study was undertaken to test the role of calcium on the gastrin mRNA production in vitro and in vivo. RIN B6 cells were cultured in medium with calcium (1.8, 3.6 and 7.2 mM) and the active form of vitamin D (calcijex). Cisplatin was added (10 μg/ml) for 12 hrs and cells were harvested for RNA from various treatment groups. Male Wistar rats were treated with cisplatin (9 mg/kg), before and after vitamin D (0.3 mg/100g/week). The rats were killed and stomach tissues excised on 1, 6, 10 and 15 days after cisplatin treatment. RNA from the stomach was analyzed using the northern blot technique. Gastrin mRNA was suppressed after cisplatin treatment both in vitro and in vivo. In vitro calcium but not vitamin D additions partially prevented the gastrin mRNA. In vivo, however, vitamin D and calcium were equally effective in preventing gastrin mRNA loss.

In-vitro- und In-vivo-Wirkung von Schilddrüsenhormon auf das Wachstum von Neuroblastomzellen

1990 ◽  
Vol 29 (03) ◽  
pp. 120-124
Author(s):  
R. P. Baum ◽  
E. Rohrbach ◽  
G. Hör ◽  
B. Kornhuber ◽  
E. Busse
Keyword(s):  
Cell Differentiation ◽  
Neuroblastoma Cells ◽  
Control Group ◽  
Initial Value ◽  
Initial Rise ◽  
Biphasic Effect ◽  
Contrast Microscopy ◽  
Morphological Sign

The effect of triiodothyronine (T3) on the differentiation of cultured neuroblastoma (NB) cells was studied after 9 days of treatment with a dose of 10-4 M/106 cells per day. Using phase contrast microscopy, 30-50% of NB cells showed formation of neurites as a morphological sign of cellular differentiation. The initial rise of the mitosis rate was followed by a plateau. Changes in cyclic nucleotide content, in the triphosphates and in the activity of the enzyme ornithine decarboxylase (ODC) were assessed in 2 human and 2 murine cell lines to serve as biochemical parameters of the cell differentiation induced by T3. Whereas the cAMP level increased significantly (3 to 7 fold compared with its initial value), the cGMP value dropped to 30 to 50% of that of the control group. ATP and GTP increased about 200%, the ODC showed a decrease of about 50%. The present studies show a biphasic effect of T3 on neuroblastoma cells: the initial rise of mitotic activity is followed by increased cell differentiation starting from day 4 of the treatment.

Radiolabeled r-Hirudin as a Measure of Thrombin Activity at, or within, the Rabbit Aorta Wall In Vitro and In Vivo

1994 ◽  
Vol 71 (04) ◽  
pp. 499-506 ◽  
Author(s):  
Mark W C Hatton ◽  
Bonnie Ross-Ouellet
Keyword(s):  
Rabbit Aorta ◽  
Balloon Injury ◽  
Recombinant Hirudin ◽  
Aorta Wall ◽  
Thrombin Activity ◽  
Before And After ◽  
The Matrix

SummaryThe behavior of 125I-labeled recombinant hirudin towards the uninjured and de-endothelialized rabbit aorta wall has been studied in vitro and in vivo to determine its usefulness as an indicator of thrombin activity associated with the aorta wall. Thrombin adsorbed to either sulfopropyl-Sephadex or heparin-Sepharose bound >95% of 125I-r-hirudin and the complex remained bound to the matrix. Binding of 125I-r-hirudin to the exposed aorta subendothelium (intima-media) in vitro was increased substantially if the tissue was pre-treated with thrombin; the quantity of l25I-r-hirudin bound to the de-endothelialized intima-media (i.e. balloon-injured in vitro) correlated positively with the quantity of bound 131I-thrombin (p <0.01). Aortas balloon-injured in vivo were measured for thrombin release from, and binding of 125I-r-hirudin to, the de-endothelialized intimal surface in vitro; 125I-r-hirudin binding correlated with the amount of active thrombin released (p <0.001). Uptake of 125I-r-hirudin by the aorta wall in vivo was proportional to the uptake of 131I-fibrinogen (as an indicator of thrombin activity) before and after balloon injury. After 30 min in the circulation, specific 125I-r-hirudin binding to the uninjured and de-endo- thelialized (at 1.5 h after injury) aorta wall was equivalent to 3.4 (± 2.5) and 25.6 (±18.1) fmol of thrombin/cm2 of intima-media, respectively. Possibly, only hirudin-accessible, glycosaminoglycan-bound thrombin is measured in this way.

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