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Sample records for adventitial vasa vasorum

  1. Hypoperfusion of the Adventitial Vasa Vasorum Develops an Abdominal Aortic Aneurysm

    PubMed Central

    Sasaki, Takeshi; Sano, Masaki; Yamamoto, Naoto; Saito, Takaaki; Inuzuka, Kazunori; Hayasaka, Takahiro; Goto-Inoue, Naoko; Sugiura, Yuki; Sato, Kohji; Kugo, Hirona; Moriyama, Tatsuya; Konno, Hiroyuki; Setou, Mitsutoshi; Unno, Naoki

    2015-01-01

    The aortic wall is perfused by the adventitial vasa vasorum (VV). Tissue hypoxia has previously been observed as a manifestation of enlarged abdominal aortic aneurysms (AAAs). We sought to determine whether hypoperfusion of the adventitial VV could develop AAAs. We created a novel animal model of adventitial VV hypoperfusion with a combination of a polyurethane catheter insertion and a suture ligation of the infrarenal abdominal aorta in rats. VV hypoperfusion caused tissue hypoxia and developed infrarenal AAA, which had similar morphological and pathological characteristics to human AAA. In human AAA tissue, the adventitial VV were stenotic in both small AAAs (30–49 mm in diameter) and in large AAAs (> 50 mm in diameter), with the sac tissue in these AAAs being ischemic and hypoxic. These results indicate that hypoperfusion of adventitial VV has critical effects on the development of infrarenal AAA. PMID:26308526

  2. Using ChromaFlo intra-vascular ultrasound (IVUS) to analyze adventitial vasa vasorum distribution: considerations and recommendations

    NASA Astrophysics Data System (ADS)

    Redwood, Abena; Holmes, David R., III; Robb, Richard

    2006-03-01

    The adventitia and outer media of large blood vessels are supplied with nutrients by microscopic blood vessels called vasa vasorum. While vasa vasorum have been implicated in a number of diseases including atherosclerosis, knowledge of their functional anatomy and specific role in these diseases has been hindered due to the small size of the vasa vasorum, and difficulty in accessing them. Micro-CT and histological methods have been used in ex-vivo animal studies of the vasa vasorum, but these techniques are limited by their inability to be used for in-vivo investigation. As such, there is very little in-vivo human data available. Intra-vascular ultrasound can acquire high-resolution anatomic images of coronary vessels. ChromaFlo IVUS has been used to identify blood flow in vessel lumens and has exciting prospect for in-vivo studies of vasa vasorum functional anatomy. In this study, ChromaFlo IVUS images of the human mid-left anterior descending coronary artery (LAD) were segmented to analyze the distribution of adventitial vasa vasorum proximal to intimal plaque. Previous animal studies suggest that formation of intimal plaque is accompanied by increased density of adventitial vasa vasorum. The data collected with ChromaFlo ultrasound is inconsistent with the current literature. While IVUS has the fidelity to acquire high-resolution US images of the coronary arteries, ChromaFlo lacks the necessary resolving power to differentiate the vasa vasorum. Further study of IVUS and other imaging methods on a large cohort will provide the basis for future in-vivo analysis of coronary disease.

  3. Quantification of Adventitial Vasa Vasorum Vascularization in Double-injury Restenotic Arteries

    PubMed Central

    Ye, Meng; Zhang, Bai-Gen; Zhang, Lan; Xie, Hui; Zhang, Hao

    2015-01-01

    Background: Accumulating evidence indicates a potential role of adventitial vasa vasorum (VV) dysfunction in the pathophysiology of restenosis. However, characterization of VV vascularization in restenotic arteries with primary lesions is still missing. In this study, we quantitatively evaluated the response of adventitial VV to vascular injury resulting from balloon angioplasty in diseased arteries. Methods: Primary atherosclerotic-like lesions were induced by the placement of an absorbable thread surrounding the carotid artery of New Zealand rabbits. Four weeks following double-injury induced that was induced by secondary balloon dilation, three-dimensional patterns of adventitial VV were reconstructed; the number, density, and endothelial surface of VV were quantified using micro-computed tomography. Histology and immunohistochemistry were performed in order to examine the development of intimal hyperplasia. Results: Results from our study suggest that double injured arteries have a greater number of VV, increased luminal surface, and an elevation in the intima/media ratio (I/M), along with an accumulation of macrophages and smooth muscle cells in the intima, as compared to sham or single injury arteries. I/M and the number of VV were positively correlated (R2 = 0.82, P < 0.001). Conclusions: Extensive adventitial VV neovascularization occurs in injured arteries after balloon angioplasty, which is associated with intimal hyperplasia. Quantitative assessment of adventitial VV response may provide insight into the basic biological process of postangioplasty restenosis. PMID:26228224

  4. Vasa vasorum and angioplasty

    SciTech Connect

    Cragg, A.H.; Einzig, S.; Rysavy, J.A.; Castaneda-Zuniga, W.R.; Borgwardt, B.; Amplatz, K.

    1983-07-01

    Interruption of flow in the vasa vasorum may lead to medial neceosis and aneurysm formation. The purpose of this study was to determine whether angioplasty produces significant alterations in the morphology or blood flow of the vasa vasorum of the dilated artery. The morphology of the canine vasa vasorum was studied before and after angioplasty; in a separate experiment vessel wall blood flow (VWBF) in canine carotid arteries was measured after angioplasty to determine whether physiologic regulation of the blood flow was disrupted by arterial dilation. No morphologic changes could be demonstrated in the vasa vasorum of the dilated artery; however, VWBF was increased. Adenosine caused a paradoxical decrease in VWBF of the dilated arterial segment while causing increased VWBF in the thoracic aorta. Angioplasty appears to produce persistent hyperemia in the dilated arterial wall. A paradoxical response to adenosine suggests that vasa vasorum in the dilated arterial segment are maximally vasodilated. This may be due to mechanical disruption of vasomotor tone or to release of vasoactive substances.

  5. Chronic adventitial inflammation, vasa vasorum expansion, and 5-lipoxygenase up-regulation in irradiated arteries from cancer survivors

    PubMed Central

    Halle, Martin; Christersdottir, Tinna; Bäck, Magnus

    2016-01-01

    Radiation-induced cardiovascular disease is an emerging problem in a steadily increasing population of survivors of cancer. However, the underlying biology is poorly described, and the late onset, which occurs several years after exposure, precludes adequate investigations in animal and cell culture models. We investigated the role of the 5-lipoxygenase (5-LO)/leukotriene pathway in radiation-induced vascular changes. Use of paired samples of irradiated arteries and nonirradiated internal control arteries from the same patient that were harvested during surgery for cancer reconstruction ≤10 yr after radiotherapy provides a unique human model of chronic radiation–induced vascular changes. Immunohistochemical stainings and perioperative inspection revealed an adventitial inflammatory response, with vasa vasorum expansion and chronic infiltration of CD68+ macrophages. These macrophages stained positive for the leukotriene-forming enzyme 5-LO. Messenger RNA levels of 5-LO and leukotriene B4 receptor 1 were increased in irradiated arterial segments compared with control vessels. These results point to targeting the 5-LO/leukotriene pathway as a therapeutic adjunct to prevent late adverse vascular effects of radiotherapy.—Halle, M., Christersdottir, T., Bäck, M. Chronic adventitial inflammation, vasa vasorum expansion, and 5-lipoxygenase up-regulation in irradiated arteries from cancer survivors. PMID:27530979

  6. Vasa vasorum in atherosclerosis and clinical significance.

    PubMed

    Xu, Junyan; Lu, Xiaotong; Shi, Guo-Ping

    2015-01-01

    Atherosclerosis is a chronic inflammatory disease that leads to several acute cardiovascular complications with poor prognosis. For decades, the role of the adventitial vasa vasorum (VV) in the initiation and progression of atherosclerosis has received broad attention. The presence of VV neovascularization precedes the apparent symptoms of clinical atherosclerosis. VV also mediates inflammatory cell infiltration, intimal thickening, intraplaque hemorrhage, and subsequent atherothrombosis that results in stroke or myocardial infarction. Intraplaque neovessels originating from VV can be immature and hence susceptible to leakage, and are thus regarded as the leading cause of intraplaque hemorrhage. Evidence supports VV as a new surrogate target of atherosclerosis evaluation and treatment. This review provides an overview into the relationship between VV and atherosclerosis, including the anatomy and function of VV, the stimuli of VV neovascularization, and the available underlying mechanisms that lead to poor prognosis. We also summarize translational researches on VV imaging modalities and potential therapies that target VV neovascularization or its stimuli.

  7. Vasa Vasorum in Atherosclerosis and Clinical Significance

    PubMed Central

    Xu, Junyan; Lu, Xiaotong; Shi, Guo-Ping

    2015-01-01

    Atherosclerosis is a chronic inflammatory disease that leads to several acute cardiovascular complications with poor prognosis. For decades, the role of the adventitial vasa vasorum (VV) in the initiation and progression of atherosclerosis has received broad attention. The presence of VV neovascularization precedes the apparent symptoms of clinical atherosclerosis. VV also mediates inflammatory cell infiltration, intimal thickening, intraplaque hemorrhage, and subsequent atherothrombosis that results in stroke or myocardial infarction. Intraplaque neovessels originating from VV can be immature and hence susceptible to leakage, and are thus regarded as the leading cause of intraplaque hemorrhage. Evidence supports VV as a new surrogate target of atherosclerosis evaluation and treatment. This review provides an overview into the relationship between VV and atherosclerosis, including the anatomy and function of VV, the stimuli of VV neovascularization, and the available underlying mechanisms that lead to poor prognosis. We also summarize translational researches on VV imaging modalities and potential therapies that target VV neovascularization or its stimuli. PMID:26006236

  8. Molecular imaging of inflammation and intraplaque vasa vasorum: A step forward to identification of vulnerable plaques?

    PubMed Central

    ten Kate, Gerrit L.; Sijbrands, Eric J. G.; Valkema, Roelf; ten Cate, Folkert J.; Feinstein, Steven B.; van der Steen, Antonius F. W.; Daemen, Mat J. A. P.

    2010-01-01

    Current developments in cardiovascular biology and imaging enable the noninvasive molecular evaluation of atherosclerotic vascular disease. Intraplaque neovascularization sprouting from the adventitial vasa vasorum has been identified as an independent predictor of intraplaque hemorrhage and plaque rupture. These intraplaque vasa vasorum result from angiogenesis, most likely under influence of hypoxic and inflammatory stimuli. Several molecular imaging techniques are currently available. Most experience has been obtained with molecular imaging using positron emission tomography and single photon emission computed tomography. Recently, the development of targeted contrast agents has allowed molecular imaging with magnetic resonance imaging, ultrasound and computed tomography. The present review discusses the use of these molecular imaging techniques to identify inflammation and intraplaque vasa vasorum to identify vulnerable atherosclerotic plaques at risk of rupture and thrombosis. The available literature on molecular imaging techniques and molecular targets associated with inflammation and angiogenesis is discussed, and the clinical applications of molecular cardiovascular imaging and the use of molecular techniques for local drug delivery are addressed. PMID:20552308

  9. Contrast-enhanced ultrasound imaging of the vasa vasorum of carotid artery plaque

    PubMed Central

    Song, Ze-Zhou; Zhang, Yan-Ming

    2015-01-01

    The vasa vasorum of carotid artery plaque is a novel marker of accurately evaluating the vulnerability of carotid artery plaque, which was associated with symptomatic cerebrovascular and cardiovascular disease. The presence of ultrasound contrast agents in carotid artery plaque represents the presence of the vasa vasorum in carotid artery plaque because the ultrasound contrast agents are strict intravascular tracers. Therefore, contrast-enhanced ultrasound (CEUS) is a novel and safe imaging modality for evaluating the vasa vasorum in carotid artery plaque. However, there are some issues that needs to be assessed to embody fully the clinical utility of the vasa vasorum in carotid artery plaque with CEUS. PMID:26120382

  10. Effects of aortic pressure and vasoactive agents on the vascular resistance of the vasa vasorum in canine isolated thoracic aorta.

    PubMed Central

    Ohhira, A; Ohhashi, T

    1992-01-01

    1. We have developed a new preparation for continuously measuring changes in vascular resistance of the vasa vasorum of the canine isolated thoracic aorta perfused at a constant flow rate with Krebs-bicarbonate solution. 2. An increase of more than 150 mmHg in aortic pressure caused a significant increase in the vascular resistance of the vasa vasorum. 3. 5-Hydroxytryptamine (5-HT), noradrenaline (NA), adrenaline and dopamine caused dose-dependent increases in the vascular resistance of the vasa vasorum. The decreasing order of potency in the vasoconstrictor responses was as follows: 5-HT >> NA = adrenaline >> dopamine. The 5-HT- and adrenaline-induced vasoconstrictor responses were inhibited by methysergide and by phentolamine plus propranolol, respectively. 4. Acetylcholine (ACh), isoprenaline (ISP), histamine (His), ATP, ADP and adenosine produced dose-related decreases in the vascular resistance of aortic vasa vasorum perfused with the Krebs solution containing 10(-5) M-NA. The decreasing order of potency in the response was as follows: ACh = ISP > His >> adenosine = ATP = ADP. The ACh-, ISP- and His-induced vasodilator responses were antagonized by atropine, propranolol and famotidine, respectively. 5. The results suggest that the preparation described is useful for studying the regulation of vascular resistance of aortic vasa vasorum and that aortic pressure and vasoactive compounds may directly regulate the vascular resistance of the vasa vasorum in canine isolated thoracic aorta. Images Fig. 2 PMID:1464829

  11. Ascending aorta of hooded seals with particular emphasis on its vasa vasorum.

    PubMed

    Blix, Arnoldus Schytte; Kuttner, Samuel; Messelt, Edward B

    2016-07-01

    The pressure-volume relationship in the ascending aorta ("windkessel") of the hooded seal was determined and the morphology of its vasa vasorum described in some detail. We found that the ascending aorta has a high compliance and can easily accommodate the entire stroke volume when the peripheral vascular resistance becomes much increased and maintain perfusion pressure during the much extended diastole and thereby reduce cardiac stroke work during diving. We also found that the 3- to 5-mm thick wall of the ascending aorta had a very elaborate vasa vasorum interna with a hitherto undescribed vascular structure that penetrates the entire vascular wall. If similar structures with similar importance for the nutrition of the wall of the vessel are found in humans, important implications for the understanding of pathological conditions, such as aneurisms, may be indicated. PMID:27122367

  12. Quantification of Vasa Vasorum Density in Multi-Slice Computed Tomographic Coronary Angiograms: Role of Computed Tomographic Image Voxel Size

    SciTech Connect

    Moritz, R.; Eaker, D; Langheinrich, A; Jorgensen, S; Bohle, R; Ritman, E

    2010-01-01

    This study is motivated by the possibility of using computed tomography (CT) to detect early coronary atherosclerosis by the increased CT values within the arterial wall resulting from vasa vasorum proliferation. Coronary arteries (n = 5) with early atherosclerotic changes were injected with Microfil and scanned (micro-CT). Noise was added to the CT projection data sets (to represent the radiation exposure of current clinical CT scanners) and then reconstructed to generate 3-dimensional images at different voxel sizes. Higher CT values were detected because of contrast agent in vasa vasorum if voxel size was less than (150 {micro}m){sup 3}. Contrast in the main lumen increased the CT values dramatically at voxels greater than (100 {micro}m){sup 3}, whereas CT values of the same specimen without contrast in the main lumen remained constant. Voxel sizes less than (200 {micro}m){sup 3} are needed to quantitate arterial wall opacification due to vasa vasorum proliferation.

  13. Atherosclerosis and Atheroma Plaque Rupture: Imaging Modalities in the Visualization of Vasa Vasorum and Atherosclerotic Plaques

    PubMed Central

    2014-01-01

    Invasive angiography has been widely accepted as the gold standard to diagnose cardiovascular pathologies. Despite its superior resolution of demonstrating atherosclerotic plaque in terms of degree of lumen stenosis, the morphological assessment for the plaque is insufficient for the analysis of plaque components, and therefore, unable to predict the risk status or vulnerability of atherosclerotic plaque. There is an increased body of evidence to show that the vasa vasorum play an important role in the initiation, progression, and complications of atherosclerotic plaque leading to major adverse cardiac events. This paper provides an overview of the evidence-based reviews of various imaging modalities with regard to their potential value for comprehensive characterization of the composition, burden, and neovascularization of atherosclerotic plaque. PMID:24688380

  14. Immortalized multipotent pericytes derived from the vasa vasorum in the injured vasculature. A cellular tool for studies of vascular remodeling and regeneration.

    PubMed

    Kabara, Maki; Kawabe, Jun-ichi; Matsuki, Motoki; Hira, Yoshiki; Minoshima, Akiho; Shimamura, Kohei; Yamauchi, Atsushi; Aonuma, Tatsuya; Nishimura, Masato; Saito, Yukihiro; Takehara, Naofumi; Hasebe, Naoyuki

    2014-12-01

    Adventitial microvessels, vasa vasorum in the vessel walls, have an active role in the vascular remodeling, although its mechanisms are still unclear. It has been reported that microvascular pericytes (PCs) possess mesenchymal plasticity. Therefore, microvessels would serve as a systemic reservoir of stem cells and contribute to the tissues remodeling. However, most aspects of the biology of multipotent PCs (mPCs), in particular of pathological microvessels are still obscure because of the lack of appropriate methods to detect and isolate these cells. In order to examine the characteristics of mPCs, we established immortalized cells residing in adventitial capillary growing at the injured vascular walls. We recently developed in vivo angiogenesis to observe adventitial microvessels using collagen-coated tube (CCT), which also can be used as an adventitial microvessel-rich tissue. By using the CCT, CD146- or NG2-positive cells were isolated from the adventitial microvessels in the injured arteries of mice harboring a temperature-sensitive SV40 T-antigen gene. Several capillary-derived endothelial cells (cECs) and PCs (cPCs) cell lines were established. cECs and cPCs maintain a number of key endothelial and PC features. Co-incubation of cPCs with cECs formed capillary-like structure in Matrigel. Three out of six cPC lines, termed capillary mPCs demonstrated both mesenchymal stem cell- and neuronal stem cell-like phenotypes, differentiating effectively into adipocytes, osteoblasts, as well as schwann cells. mPCs differentiated to ECs and PCs, and formed capillary-like structure on their own. Transplanted DsRed-expressing mPCs were resident in the capillary and muscle fibers and promoted angiogenesis and myogenesis in damaged skeletal muscle. Adventitial mPCs possess transdifferentiation potential with unique phenotypes, including the reconstitution of capillary-like structures. Their phenotype would contribute to the pathological angiogenesis associated with vascular

  15. Changes in CT angiographic opacification of porcine coronary artery wall with patchy altered flow in vasa vasorum.

    PubMed

    Moritz, Regina; Anderson, Jill L; Vercnocke, Andrew J; Wentz, Robert J; Ritman, Erik L

    2013-08-01

    To evaluate the potential of whole-body CT to detect localized areas of decreased or increased vascularity in coronary arterial walls. We used both microsphere embolization of coronary artery vasa vasorum to generate small areas of hypoperfusion and surrounding hyperperfusion of the arterial wall and diet-induced hypercholesterolemia. As a stimulus for localized angiogenesis, such as occurs in early plaque formation in the coronary arterial wall, microspheres were injected selectively into the LAD coronary artery lumens of anesthetized pigs. Fourteen pigs (acute) then had a segment of their LAD harvested during injection of contrast medium and snap-frozen for subsequent cryo-static micro-CT. An additional thirteen pigs (chronic) were allowed to recover, fed a high cholesterol diet and 3 months later were again anesthetized and a segment of the LAD artery harvested and scanned. The spatial distribution of the contrast agent within the arterial wall was measured in contiguous micro-CT images at right angles to the lumen axis with the area of wall in each cross-sectional image being approximately (0.1 mm)(3) in size. In the acute animals there were no localized areas of increased contrast around the hypoperfused embolized perfusion territories in the arterial wall, but in the chronic animals the hypoperfused areas were surrounded by increased contrast. These results suggest that CT might be able to detect localized regions of increased vascularity in the arterial wall as an indicator of early atherosclerotic stimulation of vasa vasorum proliferation. PMID:23443339

  16. Tongxinluo mitigates atherogenesis by regulating angiogenic factors and inhibiting vasa vasorum neovascularization in apolipoprotein E-deficient mice

    PubMed Central

    Ma, Lianyue; Ni, Mei; Hao, Panpan; Lu, Huixia; Yang, Xiaoyan; Xu, Xingli; Zhang, Cheng; Huang, Shanying; Zhao, Yuxia; Liu, Xiaoling; Zhang, Yun

    2016-01-01

    Vasa vasorum (VV) neovascularization contributes to atherogenesis and its expansion and distribution is correlated with intraplaque expression of angiogenic factors. The present study investigated the roles of Tongxinluo (TXL), a traditional Chinese medication, on VV proliferation and atherogenesis. In vitro, TXL pre-treatment reversed the tumor necrosis factor-a (TNF-a) induced expression of vascular endothelial growth factor A (VEGF-A) and angiopoietin-1 (ANGPT-1) but not ANGPT-2, leading to increased ratio of ANGPT-1 to ANGPT-2. Consistently, TXL treatment (at a dosage of 0.38, 0.75, 1.5 g/kg/d, respectively) decreased the expression of VEGF-A while increased that of ANGPT-1 in early atherosclerotic lesions of apolipoprotein E deficient (apoE−/−) mice. On aortic ring assay, microvessels sprouting from aortas were significantly inhibited in TXL-treated mice. Moreover, VV neovascularization in plaques was markedly reduced with TXL treatment. Histological and morphological analysis demonstrated that TXL treatment reduced plaque burden, plaque size and changed the plaque composition. These data suggest that TXL inhibits early atherogenesis through regulating angiogenic factor expression and inhibiting VV proliferation in atherosclerotic plaque. Our study shed new light on the anti-atherosclerotic effect of TXL. PMID:26908443

  17. Adventitial Vessel Growth and Progenitor Cells Activation in an Ex Vivo Culture System Mimicking Human Saphenous Vein Wall Strain after Coronary Artery Bypass Grafting

    PubMed Central

    Prandi, Francesca; Piola, Marco; Soncini, Monica; Colussi, Claudia; D’Alessandra, Yuri; Penza, Eleonora; Agrifoglio, Marco; Vinci, Maria Cristina; Polvani, Gianluca; Gaetano, Carlo; Fiore, Gianfranco Beniamino; Pesce, Maurizio

    2015-01-01

    Saphenous vein graft disease is a timely problem in coronary artery bypass grafting. Indeed, after exposure of the vein to arterial blood flow, a progressive modification in the wall begins, due to proliferation of smooth muscle cells in the intima. As a consequence, the graft progressively occludes and this leads to recurrent ischemia. In the present study we employed a novel ex vivo culture system to assess the biological effects of arterial-like pressure on the human saphenous vein structure and physiology, and to compare the results to those achieved in the presence of a constant low pressure and flow mimicking the physiologic vein perfusion. While under both conditions we found an activation of Matrix Metallo-Proteases 2/9 and of microRNAs-21/146a/221, a specific effect of the arterial-like pressure was observed. This consisted in a marked geometrical remodeling, in the suppression of Tissue Inhibitor of Metallo-Protease-1, in the enhanced expression of TGF-β1 and BMP-2 mRNAs and, finally, in the upregulation of microRNAs-138/200b/200c. In addition, the veins exposed to arterial-like pressure showed an increase in the density of the adventitial vasa vasorum and of cells co-expressing NG2, CD44 and SM22α markers in the adventitia. Cells with nuclear expression of Sox-10, a transcription factor characterizing multipotent vascular stem cells, were finally found in adventitial vessels. Our findings suggest, for the first time, a role of arterial-like wall strain in the activation of pro-pathologic pathways resulting in adventitial vessels growth, activation of vasa vasorum cells, and upregulation of specific gene products associated to vascular remodeling and inflammation. PMID:25689822

  18. Potential Role of Axonal Chemorepellent Slit2 in Modulating Adventitial Inflammation in a Rat Carotid Artery Balloon Injury Model.

    PubMed

    Liu, Dong; Xiao, Yan; Subramanian, Romesh R; Okamoto, Ei-Ichi; Wilcox, Josiah N; Anderson, Leonard; De Leon, Hector

    2016-05-01

    Leukocyte infiltration of adventitial and perivascular tissues is an early event in the development of vascular remodeling after injury. We investigated whether Slit/Robo-an axonal chemorepellent system in vertebrate and invertebrate development-is activated during the inflammatory phase that follows endothelial denudation. Using the rat carotid artery model of angioplasty, we conducted a time course analysis of mRNAs encoding Slit ligands (Slit2 and Slit3) and Robo receptors (Robo1, Robo2, and Robo4), as well as proinflammatory cell adhesion molecule (CAM) genes. Adventitial inflammatory cells were counted in immunostained arterial sections. E-selectin, vascular CAM-1, and intercellular CAM-1 were upregulated 2-3 hours after injury, followed by infiltration of neutrophils and monocytes as evidenced by real-time polymerase chain reaction, in situ hybridization, and immunohistochemistry. Slit2, Slit3, and Robo genes exhibited no expression changes at 3 hours; however, they were markedly upregulated 1 day after angioplasty. Intercellular CAM-1 expression was reduced by 50%, and the number of adventitial neutrophils decreased by >75% 1 day after angioplasty. Slit2 has been shown to be a potent chemorepelent of leukocytes, endothelial cells, and smooth muscle cells. Thus, we decided to further investigate the localization of Slit2 in injured vessels. Immunohistochemical stainings revealed the presence of Slit2 within the vessel wall and in the perivascular vasa vasorum of naive and injured arteries. Double immunohistochemical analyses showed that infiltrating monocytes expressed Slit2 in the perivascular and adventitial tissues of injured arteries 1 and 3 days postangioplasty. In addition, recombinant full-length Slit2 and Slit2-N/1118, an N-terminal fragment of Slit2, inhibited stromal cell-derived factor 1-mediated migration of circulating rat peripheral blood mononuclear cells. In summary, adventitial activation of CAM genes and neutrophil infiltration preceded

  19. Prenatal diagnosis of vasa previa.

    PubMed

    Bręborowicz, Grzegorz H; Markwitz, Wiesław; Szpera-Goździewicz, Agata; Dera-Szymanowska, Anna; Ropacka-Lesiak, Mariola; Szymański, Piotr; Kubiaczyk-Paluch, Beata

    2015-01-01

    Vasa previa is a rare condition in which unsupported by the placenta, umbilical cord blood vessels runs within the placental membranes between internal os of the cervix and presenting part of the fetus. We report an antenatal diagnostic procedure and management of a patient with low-lying placenta and velamentous cord insertion near to the internal os with two large fetal blood vessels coursing between the internal cervical os and fetal presenting part. An elective cesarean section was performed at 36 weeks gestation.

  20. A Comparative Study of the effect of Vasa Avaleha prepared with Vasa Swarasa and Vasa Kwatha in Tamaka Shwasa.

    PubMed

    Gupta, Ankit; Prajapati, P K; Choudhary, A K

    2009-01-01

    Vasa (Adhatoda vasica) is used to treat the diseases such as Shwasa, Kasa and Raktapitta in different dosage forms like Swarasa, Avaleha and Ghrita. Although the Avaleha Kalpana is not available in Brihattraya, but Gada Nigraha by Aacharya Sodhal and Bhava Prakasha have described its use in the form of Avaleha to treat the diseases of Respiratory System.The objective of this study is to compare the efficacy of two types of Vasa Avaleha prepared with either 'Swarasa' or 'Kwatha' of Vasa during their preparation. The outcomes were assessed on the basis of relief in subjective symptoms and certain hematologicalparameters.Total 35 patients were enrolled for the study. Both the groups showed highly significant results on cardinal symptoms like frequency, intensity and duration of Shwasa (dyspnoea), Kasa (coughing), Peenasa (rhinitis) with maximum percentage in Vasa Avaleha (Swarasa). The formulations also shows a insignificant decrease in haematocrit values which includes Neutrophil, Eosinophil, Lymphocyte count and TL.STL. Overall Vasa Avaleha (Swarasa) shows maximum percentage of improvement than the other group. PMID:22557317

  1. Isolation and perfusion of rat inner medullary vasa recta.

    PubMed

    Evans, Kristen K; Nawata, C Michele; Pannabecker, Thomas L

    2015-08-15

    Outer medullary isolated descending vasa recta have proven to be experimentally tractable, and consequently much has been learned about outer medullary vasa recta endothelial transport, pericyte contractile mechanisms, and tubulovascular interactions. In contrast, inner medullary vasa recta have never been isolated from any species, and therefore isolated vasa recta function has never been subjected to in vitro quantitative evaluation. As we teased out inner medullary thin limbs of Henle's loops from the Munich-Wistar rat, we found that vasa recta could be isolated using similar protocols. We isolated ∼30 inner medullary vasa recta from 23 adult male Munich-Wistar rats and prepared them for brightfield or electron microscopy, gene expression analysis by RT-PCR, or isolated tubule microperfusion. Morphological characteristics include branching and nonbranching segments exhibiting a thin endothelium, axial surface filaments radiating outward giving vessels a hairy appearance, and attached interstitial cells. Electron microscopy shows multiple cells, tight junctions, and either continuous or fenestrated endothelia. Isolated vasa recta express genes encoding the urea transporter UT-B and/or the fenestral protein PV-1, genes expressed in descending or ascending vasa recta, respectively. The transepithelial NaCl permeability (383.3 ± 60.0 × 10(-5) cm/s, mean ± SE, n = 4) was determined in isolated perfused vasa recta. Future quantitative analyses of isolated inner medullary vasa recta should provide structural and functional details important for more fully understanding fluid and solute flows through the inner medulla and their associated regulatory pathways. PMID:26062876

  2. Detection of Angiostrongylus vasorum in Red Foxes (Vulpes vulpes) from Brandenburg, Germany.

    PubMed

    Härtwig, Vera; Schulze, Christoph; Barutzki, Dieter; Schaper, Roland; Daugschies, Arwid; Dyachenko, Viktor

    2015-08-01

    Angiostrongylus (A.) vasorum is a nematode that causes angiostrongylosis in domestic and wild canids. Red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) are suspected of providing a wildlife reservoir for A. vasorum infections in pet dogs. To obtain data on the occurrence of A. vasorum in wildlife, red fox and raccoon dog carcasses (hunted or found dead) were collected from January to September 2009 in the Federal State of Brandenburg, Germany. Lung tissue samples were subjected to DNA extraction and examined for the presence of A. vasorum DNA by means of real-time PCR. A. vasorum DNA was detected in 11 out of 122 (9.0 %) lungs of red foxes and in none of the lung samples of raccoon dogs. These data suggest that red foxes are a reservoir of A. vasorum infections for pet dogs in this area.

  3. A novel form of spontaneous tool use displayed by several captive greater vasa parrots (Coracopsis vasa).

    PubMed

    Lambert, Megan L; Seed, Amanda M; Slocombe, Katie E

    2015-12-01

    Parrots are frequently cited for their sophisticated problem-solving abilities, but cases of habitual tool use among psittacines are scarce. We report the first evidence, to our knowledge, of tool use by greater vasa parrots (Coracopsis vasa). Several members of a captive population spontaneously adopted a novel tool-using technique by using pebbles and date pits either (i) to scrape on the inner surface of seashells, subsequently licking the resulting calcium powder from the tool, or (ii) as a wedge to break off smaller pieces of the shell for ingestion. Tool use occurred most frequently just prior to the breeding season, during which time numerous instances of tool transfer were also documented. These observations provide new insights into the tool-using capabilities of parrots and highlight the greater vasa parrot as a species of interest for studies of physical cognition.

  4. A novel form of spontaneous tool use displayed by several captive greater vasa parrots (Coracopsis vasa).

    PubMed

    Lambert, Megan L; Seed, Amanda M; Slocombe, Katie E

    2015-12-01

    Parrots are frequently cited for their sophisticated problem-solving abilities, but cases of habitual tool use among psittacines are scarce. We report the first evidence, to our knowledge, of tool use by greater vasa parrots (Coracopsis vasa). Several members of a captive population spontaneously adopted a novel tool-using technique by using pebbles and date pits either (i) to scrape on the inner surface of seashells, subsequently licking the resulting calcium powder from the tool, or (ii) as a wedge to break off smaller pieces of the shell for ingestion. Tool use occurred most frequently just prior to the breeding season, during which time numerous instances of tool transfer were also documented. These observations provide new insights into the tool-using capabilities of parrots and highlight the greater vasa parrot as a species of interest for studies of physical cognition. PMID:26673936

  5. The allometry of parrot BMR: seasonal data for the Greater Vasa Parrot, Coracopsis vasa, from Madagascar.

    PubMed

    Lovegrove, Barry G; Perrin, Mike R; Brown, Mark

    2011-12-01

    In this study we examined the allometry of basal metabolic rate (BMR) of 31 parrot species. Unlike previous reports, we show that parrots per se do not display BMRs that are any different to other captive-raised birds of their body size. An ordinary least squares regression fitted the data best and body mass explained 95% of the variation in BMR. There was no phylogenetic signal in the BMR data. We also provide new data for the Greater Vasa Parrot (Coracopsis vasa) of Madagascar. We tested the hypotheses that C. vasa may, because of its insular existence, display conservative energetic traits (low BMR, use of adaptive heterothermy) similar to those observed in several Malagasy mammals. However, this was not the case. C. vasa had a higher BMR than other parrots, especially during summer, when BMR was up-regulated by 50.5% and was 95.7% higher than predicted from an ordinary least squares (OLS) allometry of parrots (BMR = 0.042M (b) (0.649) , BMR in Watts, M (b) in grammes). Compared with BMR data for 94 captive-raised bird species, the winter and summer BMRs were, respectively, 45.5 and 117.8% higher than predicted by a phylogenetic generalised least squares (PGLS) allometry (BMR = 0.030M (b) (0.687) , BMR in Watts, M (b) in grammes). The summer up-regulation of BMR is the highest recorded for a bird of any size to date. We suggest that the costs of a high summer BMR may be met by the unusual cooperative breeding system of C. vasa in which groups of males feed the female and share paternity. The potential breeding benefits of a high summer BMR are unknown.

  6. A clinical review of different formulations of Vasa (Adhatoda vasica) on Tamaka Shwasa (asthma).

    PubMed

    Gupta, Ankit; Prajapati, P K

    2010-10-01

    Vasa (Adhatoda vasica Linn.) is a well known and easily available drug in almost all the seasons. Easy availability of any drug gains popularity among physicians as well as pharmaceuticals and this is the reason why almost every Kalpana of Vasa is found described in the Ayurvedika text. The different dosage forms of Vasa like Kvatha, Avaleha, Sneha, and Sandhana have been used for the treatment of Shwasa Roga. A number of research studies have been performed on different formulations of Vasa and its effect on Shwasa Roga. Therefore, a review study has been carried out on the Vasa extract, Vasa Avaleha (prepared from Svarasa and Kvatha), Vasa Ghrita, Vasarishta, and Vasakasava on Shwasa Roga, to know which formulation is better. It was found in the review that Vasa Ghana, Vasa Ghrita (1), and Vasa Avaleha have shown good results on Tamaka Shwasa. PMID:22048552

  7. An evolutionary transition of vasa regulation in echinoderms

    PubMed Central

    Juliano, Celina E.; Wessel, Gary M.

    2011-01-01

    Summary Vasa, a DEAD box helicase, is a germline marker that may also function in multipotent cells. In the embryo of the sea urchin Strongylocentrotus purpuratus, Vasa protein is posttranscriptionally enriched in the small micromere lineage, which results from two asymmetric cleavage divisions early in development. The cells of this lineage are subsequently set aside during embryogenesis for use in constructing the adult rudiment. Although this mode of indirect development is prevalent among echinoderms, early asymmetric cleavage divisions are a derived feature in this phylum. The goal of this study is to explore how vasa is regulated in key members of the phylum with respect to the evolution of the micromere and small micromere lineages. We find that although striking similarities exist between the vasa mRNA expression patterns of several sea urchins and sea stars, the time frame of enriched protein expression differs significantly. These results suggest that a conserved mechanism of vasa regulation was shifted earlier in sea urchin embryogenesis with the derivation of micromeres. These data also shed light on the phenotype of a sea urchin embryo upon removal of the Vasa-positive micromeres, which appears to revert to a basal mechanism used by extant sea stars and pencil urchins to regulate Vasa protein accumulation. Furthermore, in all echinoderms tested here, Vasa protein and/or message is enriched in the larval coelomic pouches, the site of adult rudiment formation, thus suggesting a conserved role for vasa in undifferentiated multipotent cells set aside during embryogenesis for use in juvenile development. PMID:19754712

  8. The first report of Angiostrongylus vasorum (Nematoda; Metastrongyloidea) in Poland, in red foxes (Vulpes vulpes).

    PubMed

    Demiaszkiewicz, Aleksander W; Pyziel, Anna M; Kuligowska, Izabela; Lachowicz, Jacek

    2014-10-01

    Angiostrongylus vasorum belongs to the superfamily of Metastrongyloidea. This nematode occurs in foxes, dogs and other predators. The Nematode A. vasorum place themselves in the pulmonary artery and its branches, and in the right ventricle and atrium of the heart. Numerous species of land snails are the intermediate hosts of the parasite. In 2013, lungs and hearts of 76 foxes shot in the Forest District Głęboki Bród in Augustowska Primeval Forest were parasitologically necropsied. Four of the examined foxes were infected with the nematode A. vasorum, a prevalence of 5.2%. In one fox pericardium there were 6 male and 6 female nematodes. In the remaining three foxes nematodes were localized in the pulmonary artery. In two foxes 2 specimens of nematodes were detected (male and female, and two females) while 1 female was detected in the other fox. This is the first report of the presence of the nematode A. vasorum in fox in Poland.

  9. Molecular identification of novel intermediate host species of Angiostrongylus vasorum in Greater London.

    PubMed

    Patel, Zainab; Gill, A Christina; Fox, Mark T; Hermosilla, Carlos; Backeljau, Thierry; Breugelmans, Karin; Keevash, Esther; McEwan, Claudia; Aghazadeh, Mahdis; Elson-Riggins, Jocelyn G

    2014-12-01

    Angiostrongylus vasorum is a parasitic nematode that can cause serious and potentially fatal disease in dogs and other canids. The aim of this study was to determine the intermediate slug species infected in nature by sampling sites in Greater London and Hertfordshire located within a known hyperendemic region. Overall, A. vasorum larvae were recovered from 6/381 slugs (1.6%) by tissue digestion, and their identity was confirmed by PCR. Infected slugs originated from three different sites in the Greater London area: one in Waltham Forest and two in Bromley. Slugs parasitised by A. vasorum were identified by a combination of external morphological characteristics and molecular techniques and belonged to three different families: the Arionidae, the Milacidae and the Limacidae. This includes two new host records for the parasite: Arion distinctus and Tandonia sowerbyi. This is the first record of A. vasorum in the family Milacidae, indicating that the parasite has a broader intermediate host range than previously recognised.

  10. Zebrafish vasa is required for germ-cell differentiation and maintenance.

    PubMed

    Hartung, Odelya; Forbes, Meredyth M; Marlow, Florence L

    2014-10-01

    Vasa is a universal marker of the germ line in animals, yet mutations disrupting vasa cause sexually dimorphic infertility, with impaired development of the ovary in some animals and the testis in others. The basis for this sexually dimorphic requirement for Vasa is not clear; in most animals examined, both the male and female gonad express vasa throughout the life of the germ line. Here we characterized a loss-of-function mutation disrupting zebrafish vasa. We show that maternally provided Vasa is stable through the first ten days of development in zebrafish, and thus likely fulfills any early roles for Vasa during germ-line specification, migration, survival, and maintenance. Although zygotic Vasa is not essential for the development of juvenile gonads, vasa mutants develop exclusively as sterile males. Furthermore, phenotypes of vasa;p53 compound mutants are indistinguishable from those of vasa mutants, therefore the failure of vasa mutants to differentiate as females and to support germ-cell development in the testis is not due to p53-mediated apoptosis. Instead, we found that failure to progress beyond the pachytene stage of meiosis causes the loss of germ-line stem cells, leaving empty somatic tubules. Our studies provide insight into the function of zebrafish vasa during female meiosis, differentiation, and maintenance of germ-line stem cells.

  11. A case report of vasa previa incidentally discovered.

    PubMed

    Saghir, Salahiddine; Kouach, Jaouad; Agadr, Aomar

    2015-01-01

    Vasa previa is a rare but clinically important obstetrical complication that can be associated with a low-lying placenta or placenta previa. We aim to present one case of vasa previa diagnosed during the placenta examination after the caesarean indicated for triple uterus scar. A 26-year-old female was referred to our hospital at 30 weeks of gestation to provide a scheduled caesarean. Trans-abdominal ultrasound was performed; the placenta was positioned in the posterior side of the fundus. Fetal growth was found to be appropriate for gestational age. A healthy male infant weighing was successfully delivered via cesarean section at 38 weeks of gestation. This operation helped to prevent complications due to acute fetal bleeding. The identification and exclusion of vasa previa using trans-vaginal ultrasound are essential to ensure appropriate and timely treatment.

  12. Essential elements for translation: the germline factor Vasa functions broadly in somatic cells

    PubMed Central

    Yajima, Mamiko; Wessel, Gary M.

    2015-01-01

    ABSTRACT Vasa is a conserved RNA-helicase found in the germ lines of all metazoans tested. Whereas Vasa presence is often indicated as a metric for germline determination in animals, it is also expressed in stem cells of diverse origin. Recent research suggests, however, that Vasa has a much broader function, including a significant role in cell cycle regulation. Results herein indicate that Vasa is utilized widely, and often induced transiently, during development in diverse somatic cells and adult precursor tissues. We identified that Vasa in the sea urchin is essential for: (1) general mRNA translation during embryogenesis, (2) developmental re-programming upon manipulations to the embryo and (3) larval wound healing. We also learned that Vasa interacted with mRNAs in the perinuclear area and at the spindle in an Importin-dependent manner during cell cycle progression. These results suggest that, when present, Vasa functions are essential to contributing to developmental regulation. PMID:25977366

  13. Angiostrongylus vasorum infection in red foxes (Vulpes vulpes) in southern Italy.

    PubMed

    Santoro, Mario; D'Alessio, Nicola; Di Prisco, Francesca; Neola, Benedetto; Restucci, Brunella; Pagano, Teresa B; Veneziano, Vincenzo

    2015-06-01

    Angiostrongylus vasorum (Nematoda: Angiostrongylidae) infection was detected at post-mortem examination in the pulmonary arteries and hearts of 34/102 (33,3%) of red foxes (Vulpes vulpes) from the Campania Region in southern Italy. Pathological changes consisted of granulomatous interstitial pneumonia caused by larvae and intravascular pulmonary adult nematodes. These changes confirm that angiostrongylosis infection in red foxes has a mainly chronic course, in which the infected host may disperse parasite larvae in the environment over its lifetime. Results suggest that the life cycle of A. vasorum is well established in the red fox in the Campania Region representing a potential infection risk for dogs.

  14. Angiostrongylus vasorum Causing Severe Granulomatous Hepatitis with Concurrent Multiple Acquired PSS.

    PubMed

    Cook, Simon; Priestnall, Simon L; Blake, Damer; Meeson, Richard L

    2015-01-01

    A 14 mo old female Jack Russell terrier presented with a 12 hr history of vomiting and inappetence. She was subsequently diagnosed with multiple acquired portosystemic shunts during an exploratory celiotomy. Gross and histopathological hepatic abnormalities were consistent with chronic disease, including features suggestive of portal hypertension that was potentially caused by migrating and resident Angiostrongylus vasorum larvae. Fecal analysis and polymerase chain reaction of hepatic tissue confirmed the presence of Angiostrongylus vasorum . The dog recovered clinically following empirical treatment and supportive care. A lack of parasite burden was confirmed 9 wk postdiagnosis; however, serum biochemical analysis at that time was suggestive of ongoing hepatic dysfunction.

  15. [Adventitial cyst of the aorta].

    PubMed

    Kitzis, M; Assens, P; Couffinhal, J C; Bourgeois, P; Weiss, A M; Remond, P; Andreassian, B

    1983-11-10

    The authors report a case of adventitial cyst of the abdominal aorta. This would seem to be the first time that a cyst has been reported in the literature with this localization. A 54 year old female patient was treated surgically for a suspected aneurysm of the abdominal aorta. Her antecedents included syphilis treated with bismuth. The condition began with a lumbago with no clearly defined etiology. An abdominal X-ray without preparation visualized a calcified abdominal tumor in a retroperitoneal position, apparently attached to the aorta. Arteriography showed that the tumor was excluded from the circulation. Surgical exploration of the aorta demonstrated a cystic tumor, which was resected. A review of the literature concerning the pathology of arterial cysts shows no previous cases of aortic cyst. On the other hand, references to identical anomalies of the iliac, femoral, popliteal and radial arteries provide grounds for discussion of the etiology of this aortic disease. The macroscopic data and histological studies suggest a phenomenon of cystic degeneration rather than a constitutional anomaly of the artery, or, in this particular case, an anomaly connected with syphilitic lesions.

  16. Post-translational regulation by gustavus contributes to selective Vasa protein accumulation in multipotent cells during embryogenesis

    PubMed Central

    Gustafson, Eric A.; Yajima, Mamiko; Juliano, Celina E.; Wessel, Gary M.

    2010-01-01

    Vasa is a broadly conserved DEAD-box RNA helicase associated with germ line development and is expressed in multipotent cells in many animals. During embryonic development of the sea urchin Strongylocentrotus purpuratus, Vasa protein is enriched in the small micromeres despite a uniform distribution of vasa transcript. Here we show that the Vasa coding region is sufficient for its selective enrichment and find that gustavus, the B30.2/SPRY and SOCS box domain gene, contributes to this phenomenon. In vitro binding analyses show that Gustavus binds the N-terminal and DEAD-box portions of Vasa protein independently. A knockdown of Gustavus protein reduces both Vasa protein abundance and its propensity for accumulation in the small micromeres, whereas overexpression of the Vasa-interacting domain of Gustavus (GusΔSOCS) results in Vasa protein accumulation throughout the embryo. We propose that Gustavus has a conserved, positive regulatory role in Vasa protein accumulation during embryonic development. PMID:21035437

  17. Post-translational regulation by gustavus contributes to selective Vasa protein accumulation in multipotent cells during embryogenesis.

    PubMed

    Gustafson, Eric A; Yajima, Mamiko; Juliano, Celina E; Wessel, Gary M

    2011-01-15

    Vasa is a broadly conserved DEAD-box RNA helicase associated with germ line development and is expressed in multipotent cells in many animals. During embryonic development of the sea urchin Strongylocentrotus purpuratus, Vasa protein is enriched in the small micromeres despite a uniform distribution of vasa transcript. Here we show that the Vasa coding region is sufficient for its selective enrichment and find that gustavus, the B30.2/SPRY and SOCS box domain gene, contributes to this phenomenon. In vitro binding analyses show that Gustavus binds the N-terminal and DEAD-box portions of Vasa protein independently. A knockdown of Gustavus protein reduces both Vasa protein abundance and its propensity for accumulation in the small micromeres, whereas overexpression of the Vasa-interacting domain of Gustavus (GusΔSOCS) results in Vasa protein accumulation throughout the embryo. We propose that Gustavus has a conserved, positive regulatory role in Vasa protein accumulation during embryonic development. PMID:21035437

  18. Larval development of Angiostrongylus vasorum in the land snail Helix aspersa.

    PubMed

    Di Cesare, Angela; Crisi, Paolo Emidio; Bartolini, Roberto; Iorio, Raffaella; Talone, Tonino; Filippi, Laura; Traversa, Donato

    2015-10-01

    The metastrongyloid nematode Angiostrongylus vasorum affects the heart and pulmonary arteries of dogs and wild animals. Over the recent years, dog angiostrongylosis has gained great attention in the veterinary community for the expansion of its geographic range and for a rise in the number of clinical cases. Global warming, changes in phenology of mollusc intermediate hosts and movements of wild reservoirs have been evocated in the spreading of mollusc-borne parasites, including A. vasorum. The land snail Helix aspersa, a vector of other respiratory metastrongyloids, is endemic in most regions of the World, where it is a pest outside its native Mediterranean range. In the present study, the susceptibility and suitability of H. aspersa as an intermediate host of A. vasorum were investigated along with the characteristics of larval recovery and development following two different ways of inoculation, i.e. experimental (group A) vs natural infection (group B). After infections, the snails were kept at environmental conditions for 2 months. Five snails from groups A and B were randomly selected, digested and examined at 15-day intervals for 2 months. L1s, L2s and L3s were microscopically identified based on key morphological and morphometric characteristics and their identity was genetically confirmed. The results showed that A. vasorum may reach the infective stage in H. aspersa and that uptake of larvae and parasitic burden within the snails depend on the grazing capability of the molluscs. Biological and epidemiological implications are discussed.

  19. Epidemiological survey of Angiostrongylus vasorum in dogs and slugs around a new endemic focus in Scotland.

    PubMed

    Helm, J; Roberts, L; Jefferies, R; Shaw, S E; Morgan, E R

    2015-07-11

    The nematode parasite Angiostrongylus vasorum is an increasingly important cause of respiratory and other diseases in dogs. Geographical spread from previously limited endemic foci has occurred rapidly. This paper investigates parasite epidemiology around the location of the first reported case in Scotland in 2009: by detection of A vasorum-specific DNA in gastropod intermediate hosts, and in dogs circulating DNA and specific antibodies, and first stage larvae in faeces. Overall prevalence in gastropods was 6.7 per cent (16/240), with parasite DNA found in slugs in the Arion ater and Arion hortensis species aggregates and the snail Helix aspersa (syn. Cornu aspersum). Of 60 dogs presenting with clinical signs compatible with angiostrongylosis, none tested positive using PCR on peripheral blood or Baermann test on faeces, and none of 35 tested for circulating anti-A vasorum antibodies were positive. PCR prevalence in gastropods was highest (11 per cent) in the park frequented by the canine angiostrongylosis index case. Molecular survey for infection in gastropods is a potentially informative and efficient method for characterising the distribution of A vasorum and therefore local risk of canine infection. However, there appears to be a complex relationship between prevalence in gastropods and emergence of canine clinical disease, which requires further work to advance understanding of parasite transmission and geographical disease spread.

  20. First report of a fatal autochthonous canine Angiostrongylus vasorum infection in Belgium.

    PubMed

    Jolly, Sandra; Poncelet, Luc; Lempereur, Laetitia; Caron, Yannick; Bayrou, Calixte; Cassart, Dominique; Grimm, Felix; Losson, Bertrand

    2015-02-01

    Canine angiostrongylosis is considered as an emergent disease in Europe and Canada. A fatal case of Angiostrongylus vasorum infection is described in a four and a half month old puppy born in Belgium. The dog was presented with marked neurological disorders, body weight loss, a profound weakness and mild respiratory signs. The dog was given antibiotics and mucolytic compounds with very little improvement and consequently was referred to a specialist for additional examinations. As the general condition of the dog was rapidly declining, the animal was euthanized shortly after on its owners' request and a necropsy was carried out. Extensive gross pulmonary lesions were observed and histopathological examination revealed the presence of numerous larvae with morphology compatible with A. vasorum. Larvae were also found in the product of a bronchoalveolar lavage but fecal material was not examined. The presence of A. vasorum circulating serum antigen was demonstrated through ELISA; additionally an A. vasorum specific PCR was performed on brain material and yielded a positive result. This case confirms that the clinical diagnosis of canine angiostrongylosis can be very challenging especially when respiratory signs are absent or very mild such in the present case. This is the first reported case of canine angiostrongylosis in Belgium.

  1. Severe pulmonary arterial hypertension due to Angiostrongylosus vasorum in a dog.

    PubMed

    Nicolle, Audrey P; Chetboul, Valérie; Tessier-Vetzel, Dominique; Carlos Sampedrano, Carolina; Aletti, Edouard; Pouchelon, Jean-Louis

    2006-08-01

    A dog was presented with a history of dyspnea, coughing, and ascites. Angiostrongylosis and severe pulmonary arterial hypertension (PAH) were found, as well as a marked discordance between the electrical and mechanical events of the heart. Pulmonary arterial hypertension related to Angiostrongylus vasorum has rarely been reported.

  2. Larval development of Angiostrongylus vasorum in the land snail Helix aspersa.

    PubMed

    Di Cesare, Angela; Crisi, Paolo Emidio; Bartolini, Roberto; Iorio, Raffaella; Talone, Tonino; Filippi, Laura; Traversa, Donato

    2015-10-01

    The metastrongyloid nematode Angiostrongylus vasorum affects the heart and pulmonary arteries of dogs and wild animals. Over the recent years, dog angiostrongylosis has gained great attention in the veterinary community for the expansion of its geographic range and for a rise in the number of clinical cases. Global warming, changes in phenology of mollusc intermediate hosts and movements of wild reservoirs have been evocated in the spreading of mollusc-borne parasites, including A. vasorum. The land snail Helix aspersa, a vector of other respiratory metastrongyloids, is endemic in most regions of the World, where it is a pest outside its native Mediterranean range. In the present study, the susceptibility and suitability of H. aspersa as an intermediate host of A. vasorum were investigated along with the characteristics of larval recovery and development following two different ways of inoculation, i.e. experimental (group A) vs natural infection (group B). After infections, the snails were kept at environmental conditions for 2 months. Five snails from groups A and B were randomly selected, digested and examined at 15-day intervals for 2 months. L1s, L2s and L3s were microscopically identified based on key morphological and morphometric characteristics and their identity was genetically confirmed. The results showed that A. vasorum may reach the infective stage in H. aspersa and that uptake of larvae and parasitic burden within the snails depend on the grazing capability of the molluscs. Biological and epidemiological implications are discussed. PMID:26122991

  3. [Research progress of adventitious respiratory sound signal processing].

    PubMed

    Li, Zhenzhen; Wu, Xiaoming

    2013-10-01

    Adventitious respiratory sound signal processing has been an important researching topic in the field of computerized respiratory sound analysis system. In recent years, new progress has been achieved in adventitious respiratory sound signal analysis due to the applications of techniques of non-stationary random signal processing. Algorithm progress of adventitious respiratory sound detections is discussed in detail in this paper. Then the state of art of adventitious respiratory sound analysis is reviewed, and development directions of next phase are pointed out.

  4. Cloning and expression analysis of vasa during the reproductive cycle of Korean rockfish, Sebastes schlegeli

    NASA Astrophysics Data System (ADS)

    Mu, Weijie; Wen, Haishen; He, Feng; Li, Jifang; Liu, Miao; Ma, Ruiqin; Zhang, Yuanqing; Hu, Jian; Qi, Baoxia

    2013-03-01

    Vasa, which is a conserved member of the DEAD-box protein family, plays an indispensable role in primordial germ cell proliferation. However, the expression of vasa gene during the reproductive cycle in ovoviviparous fish has not been documented. In this study, the full-length sequence of vasa was obtained from the ovary of Korean rockfish ( Sebastes schlegeli) using reverse transcription-PCR and rapid amplification of cDNA ends. The Vasa with a mature protein of 650 amino acids showed greatest homology (84%) with giant gourami ( Osphronemus goramy) and Pacific bluefin tuna ( Thunnus orientalis). The expression of vasa mRNA in Korean rockfish was detected in gonads only, suggesting its specific role in gonadal development. In addition, seasonal changes in the vasa expression levels were examined in gonads by quantitative real-time PCR. The vasa transcript levels in adult testis were found higher during spermatogenesis than during spermiation. The vasa transcript levels remained relatively high at the early ovary stage but declined during ovary maturation in adult female fish. These results suggest that the vasa gene play an important role in spermatogenesis and early oogenesis during the reproductive cycle of Korean rockfish.

  5. Preliminary characterization and expression of Vasa-like gene in Schistosoma japonicum.

    PubMed

    Diao, Yujie; Hua, Mengqing; Shao, Yanjing; Huang, Wei; Liu, Miao; Ren, Cuiping; Ji, Yongsheng; Chen, Jianmin; Shen, Jijia

    2015-07-01

    The Vasa gene is a vital germline marker to study the origin and development of germ cells and gonads in many organisms. Until now, little information was available about the characteristics of the Vasa gene in Schistosoma japonicum (S. japonicum). In this study, we cloned the open reading frame (ORF) of the S. japonicum Vasa-like gene (Sj-Vasa). The expression pattern and tissue localization of Sj-Vasa were also analyzed. Our results showed that Sj-Vasa shared the general feature of DEAD-box family member proteins. Sj-Vasa was transcribed and expressed throughout the S. japonicum life cycle with transcription exhibiting high levels at day 24 in both male and female worms, and the expression level in the female was always higher than that in the male. Sj-Vasa protein was localized in a variety of tissues of adult schistosomes, including the gonads (ovary, vitellarium, and testes), the subtegument, and some cells of the parenchyma. To our knowledge, this is the first report of preliminary characterization and expression of the Vasa-like gene that may play an important role in the development of the worm, especially in reproductive organs of S. japonicum. PMID:25899325

  6. [Construction of goat germ cell specific reporting system pVASA-EGFP].

    PubMed

    Yan, Guangyao; Li, Peizhen; Ren, Caifang; Wang, Feng; Zhang, Yanli

    2015-09-01

    To monitor the trans-differentiation from adult stem cells to germ cells, we analyzed the vasa expression of goat testicular tissues in different ages and constructed the germ cell specific reporting vector pVASA-EGFP. The expression of vasa was verified by RT-PCR and immunofluorescence. The vector pVASA-EGFP was constructed by molecular technology, then transfected into goat bone mesenchymal stem cells (BMSCs) by Lipofectamine 2000. Moreover, we observed the expression of the vector through green fluorescent protein (GFP). Immunofluorescence results show that Vasa was expressed in all groups of goat testicular tissues, RT-PCR results show that the levels of vasa mRNA in 3-month group and 10-month group were significantly higher than that in 10-day group. Sequencing and restriction enzyme results show that the vector was successfully constructed. After transfection and RA treatment, GFP expression was observed, which proved the validity of our reporting system. All the results proved that vasa was expressed in different ages in goat testicular tissues, and the vector pVASA-EGFP is efficient in monitoring the trans-differentiation in vitro, which paves the way for further characterization and screening of the trans-differentiation of goat BMSCs.

  7. [Princess Anna Vasa--her fascinating life story and skeleton].

    PubMed

    During, Ebba

    2005-01-01

    The Princess Anna Vasa was born in Sweden in 1568 and spent her first 19 years there. She was the daughter of the Swedish king Johan III and his wife, the Polish Royal Princess Katarina Jagellonica. She was brought up as a Catholic but converted to be a Protestant already in 1583 and remained a fervent Protestant to the end of her life. She was an exceptionally intelligent and extensively educated woman. When her brother became king, Sigismund III of Poland, she accompanied him there. She exerted great influence on Sigismund who was brought up to be a Catholic. She was persistent in her religion, yet working for religious liberty. "The Swedish Princess" was also named "the Queen of Polish Botany". She was never married and she died 57 years old in 1625. For religious reasons her body had to wait 11 years for a funeral of royal standing. The funeral took place in 1636 in St Mary's Church in Torun, Poland. During restoration work at the church in April 1994, Anna Vasa's skeleton was removed from the tomb, and an antropological investigation in order to establish her identity was carried by Dr Andrzej Florkowski at the Dept of Anthrop, Nicholas Copernicus University of Torun. I was invited to Torun to examine her remains in May 1995. The skeleton was in a rather good state of preservation. However, her grave had been plundered at least twice. Her skeleton lacked the right forearm and hand, probably as the result of the pillage of her rings and bracelets. Some other bones and teeth were also missing. At our ocular examination the skeleton revealed a number of anatomical deformations and pathological changes. A conventional radiography and CT of Anna Vasa's skeletal remains was later carried out in 1995 by M. Grzegorzewski, Z. Boron and W. Lasek at the Dept of Radiology, Med. Acad. of Bydgoszcz, Polen. A DNA-analysis was carried out by Dr Anders Götherström at the Archaeol. Res. Lab., Stockholm Univ. An odontological and radiological study was performed by Dr Sigrid I

  8. Vasa genes: Emerging roles in the germ line and in multipotent cells

    PubMed Central

    Gustafson, Eric A.; Wessel, Gary M.

    2011-01-01

    Sexually reproducing metazoans establish a cell lineage during development that is ultimately dedicated to gamete production. Work in a variety of animals suggests that a group of conserved molecular determinants function in this germ line maintenance and function. The most universal of these genes are vasa and vasa-like DEAD box RNA helicase genes. However, recent evidence indicates that vasa genes also function in other cell types, distinct from the germ line. Here we evaluate our current understanding of vasa function and its regulation during development, addressing vasa’s emerging role in multipotent cells. We also explore the evolutionary diversification of the amino-terminal domain of this gene and how this impacts the association of vasa with nuage-like perinuclear structures. PMID:20586054

  9. Nonsteroidal anti-inflammatory drugs alter vasa recta diameter via pericytes.

    PubMed

    Kennedy-Lydon, Teresa; Crawford, Carol; Wildman, Scott S; Peppiatt-Wildman, Claire M

    2015-10-01

    We have previously shown that vasa recta pericytes are known to dilate vasa recta capillaries in the presence of PGE2 and contract vasa recta capillaries when endogenous production of PGE2 is inhibited by the nonselective nonsteroidal anti-inflammatory drug (NSAID) indomethacin. In the present study, we used a live rat kidney slice model to build on these initial observations and provide novel data that demonstrate that nonselective, cyclooxygenase-1-selective, and cyclooxygenase -2-selective NSAIDs act via medullary pericytes to elicit a reduction of vasa recta diameter. Real-time images of in situ vasa recta were recorded, and vasa recta diameters at pericyte and nonpericyte sites were measured offline. PGE2 and epoprostenol (a prostacyclin analog) evoked dilation of vasa recta specifically at pericyte sites, and PGE2 significantly attenuated pericyte-mediated constriction of vasa recta evoked by both endothelin-1 and ANG II. NSAIDs (indomethacin > SC-560 > celecoxib > meloxicam) evoked significantly greater constriction of vasa recta capillaries at pericyte sites than at nonpericyte sites, and indomethacin significantly attenuated the pericyte-mediated vasodilation of vasa recta evoked by PGE2, epoprostenol, bradykinin, and S-nitroso-N-acetyl-l-penicillamine. Moreover, a reduction in PGE2 was measured using an enzyme immune assay after superfusion of kidney slices with indomethacin. In addition, immunohistochemical techniques were used to demonstrate the population of EP receptors in the medulla. Collectively, these data demonstrate that pericytes are sensitive to changes in PGE2 concentration and may serve as the primary mechanism underlying NSAID-associated renal injury and/or further compound-associated tubular damage. PMID:26202223

  10. Comparison of three methods for the detection of Angiostrongylus vasorum in the final host.

    PubMed

    Houpin, E; McCarthy, G; Ferrand, M; De Waal, T; O'Neill, E J; Zintl, A

    2016-04-15

    Angiostrongylosis is potentially fatal parasitic nematode infection affecting dogs which can be difficult to diagnose. In recent years several microscopical, serological and molecular detection methods have been developed, however there are few studies that have compared the relative performance of these methods. Screening necropsy material from an opportunistic sample of 140 foxes (82 of which were considered to be infected with Angiostrongylus vasorum), indicated sensitivities of 84.1% for dissection and visual examination of plucks, 69.5% for nested PCR of an 18S rRNA fragment and 76.8% for a canine A. vasorum antigen detection test (IDEXX Angio Detect) of tissue fluid samples respectively. Agreement between the tests ranged from 45.6 to 79.7%. A novel nested PCR-RFLP for the detection and identification of canid lungworm spp. is described.

  11. Rupture of the thoracic aorta associated with experimental Angiostrongylus vasorum infection in a dog.

    PubMed

    Mozzer, L R; Lima, W S

    2012-05-01

    This note describes the sudden death of a dog by the rupture of the thoracic aorta caused by the presence of Angiostrongylus vasorum. A female mongrel canine with a history of weight loss and exhaustion died two hours after clinical examination. At necropsy, performed one hour after death, showed the presence of clotted blood in the thoracic cavity. Haemothorax was diagnosed. The thoracic aorta wall was thin, congested and an abnormal hole in the wall was detected approximately 0.5 cm from the entrance to the diaphragm. From clotted blood collected from the thoracic cavity, 224 first stage larvae (L1) and 15 adults of Angiostrongylus vasorum were recovered alive. Also, from a blood clot found in the aorta, four adult females and 47 L1 larvae were recovered alive. Possibly, this parasite was responsible for the aortic rupture and death of the animal.

  12. Strigolactones suppress adventitious rooting in Arabidopsis and pea.

    PubMed

    Rasmussen, Amanda; Mason, Michael Glenn; De Cuyper, Carolien; Brewer, Philip B; Herold, Silvia; Agusti, Javier; Geelen, Danny; Greb, Thomas; Goormachtig, Sofie; Beeckman, Tom; Beveridge, Christine Anne

    2012-04-01

    Adventitious root formation is essential for the propagation of many commercially important plant species and involves the formation of roots from nonroot tissues such as stems or leaves. Here, we demonstrate that the plant hormone strigolactone suppresses adventitious root formation in Arabidopsis (Arabidopsis thaliana) and pea (Pisum sativum). Strigolactone-deficient and response mutants of both species have enhanced adventitious rooting. CYCLIN B1 expression, an early marker for the initiation of adventitious root primordia in Arabidopsis, is enhanced in more axillary growth2 (max2), a strigolactone response mutant, suggesting that strigolactones restrain the number of adventitious roots by inhibiting the very first formative divisions of the founder cells. Strigolactones and cytokinins appear to act independently to suppress adventitious rooting, as cytokinin mutants are strigolactone responsive and strigolactone mutants are cytokinin responsive. In contrast, the interaction between the strigolactone and auxin signaling pathways in regulating adventitious rooting appears to be more complex. Strigolactone can at least partially revert the stimulatory effect of auxin on adventitious rooting, and auxin can further increase the number of adventitious roots in max mutants. We present a model depicting the interaction of strigolactones, cytokinins, and auxin in regulating adventitious root formation. PMID:22323776

  13. The development of Angiostrongylus vasorum (Baillet, 1866) in the freshwater snail Pomacea canaliculata (Lamarck, 1822).

    PubMed

    Mozzer, L R; Coaglio, A L; Dracz, R M; Ribeiro, V M A; Lima, W S

    2015-11-01

    Angiostrongylus vasorum is a parasitic nematode that infects the heart and pulmonary artery and its branches of domestic and wild canids. The parasite can use several species of terrestrial and aquatic molluscs as intermediate hosts, although susceptibility varies. Pomacea canaliculata is a mollusc found in lakes, swamps and rivers in South America. In this study, we evaluated the susceptibility, parasite growth, oviposition and larval development of 282 P. canaliculata infected with 500 A. vasorum first-instar larvae (L1). From day 5 post-infection (pi) to day 30 pi, seven specimens per day were sacrificed to recover the larval instars. We compared 50 egg masses from infected and uninfected molluscs to determine the number of eggs per clutch, the hatching rate and the growth of the molluscs. The percentage of recovered larvae ranged from 39.17% to 67.5%. First-stage larvae (L1) were found until day 19 pi, second-stage larvae (L2) were found from days 11 to 25 pi, and third-stage larvae (L3) were recovered only after day 19 pi. Infected snails exhibited the most eggs during spawning, although the rate of hatching and shell size were lower in the infected snails compared with controls. This is the first report of an experimental infection of P. canaliculata with A. vasorum, and the results confirm the non-specificity of the nematode in relation to the intermediate host and indicate the importance of epidemiological surveys of this parasite and mollusc.

  14. Pneumonia from Angiostrongylus vasorum infection in a red panda (Ailurus fulgens fulgens).

    PubMed

    Patterson-Kane, Janet C; Gibbons, Lynda M; Jefferies, Ryan; Morgan, Eric R; Wenzlow, Nanny; Redrobe, Sharon P

    2009-03-01

    A 9-year-old, male, captive red panda (Ailurus fulgens fulgens) in an urban zoo in the United Kingdom presented with respiratory distress and weight loss. The animal was euthanatized, and a postmortem examination was performed. The lungs were diffusely consolidated with extensive mineralization. Microscopically, there was extensive obliteration of normal pulmonary architecture by sheets and coalescing nodules of partially mineralized fibrous tissue and granulomatous inflammation centered on large numbers of nematode larvae and eggs. First stage nematode larvae were isolated from lung tissue and were characterized as Angiostrongylus vasorum on the basis of their morphology and sequencing of the 18S ribosomal RNA gene and the entire second internal transcribed spacer. Although A. vasorum has previously been reported in red pandas in a zoological collection in Denmark, this study is the first reported case in the United Kingdom and occurs against a background of geographical spread and increased incidence of disease in domestic and wild canids. Angiostrongylus vasorum should be considered a differential diagnosis for respiratory disease in the red panda and taken into account when planning parasite and pest control programs for zoological collections.

  15. First report of Angiostrongylus vasorum and Hepatozoon from a red fox (Vulpes vulpes) from West Virginia, USA.

    PubMed

    Kistler, Whitney M; Brown, Justin D; Allison, Andrew B; Nemeth, Nicole M; Yabsley, Michael J

    2014-02-24

    Angiostrongylus vasorum was identified in the lungs of a red fox (Vulpes vulpes) from West Virginia, United States (US), indicating a new geographical location for this metastrongylid nematode. The fox was euthanized and submitted for necropsy after displaying erratic behavior. We did not detect rabies virus or canine distemper virus from the fox. We observed bronchopneumonia associated with A. vasorum infection disseminated in both lungs. In addition, protozoal meronts were observed in the liver, spleen, and mesenteric lymph node, and were identified as Hepatozoon canis. Lymphoid depletion was also observed in the spleen and mesenteric lymph node. In addition to A. vasorum and H. canis infections, Eucoleus aerophilus eggs and adult worms were observed in the lungs of the fox. Severe lesions associated with A. vasorum infection were observed in the lungs and these were determined to be the likely cause of morbidity; however, synergistic effects among the multiple infections detected in this fox cannot be ruled out. This is the first report of an autochthonous A. vasorum infection in the US and from outside of Newfoundland Canada, the only place in North America where the parasite is known to be endemic. Additionally, this is the first report of a H. canis infection in a red fox from the US. PMID:24412356

  16. Abnormal Placentation: Placenta Previa, Vasa Previa, and Placenta Accreta.

    PubMed

    Silver, Robert M

    2015-09-01

    Placental disorders such as placenta previa, placenta accreta, and vasa previa are all associated with vaginal bleeding in the second half of pregnancy. They are also important causes of serious fetal and maternal morbidity and even mortality. Moreover, the rates of previa and accreta are increasing, probably as a result of increasing rates of cesarean delivery, maternal age, and assisted reproductive technology. The routine use of obstetric ultrasonography as well as improving ultrasonographic technology allows for the antenatal diagnosis of these conditions. In turn, antenatal diagnosis facilitates optimal obstetric management. This review emphasizes an evidence-based approach to the clinical management of pregnancies with these conditions as well as highlights important knowledge gaps.

  17. Insights into the immuno-molecular biology of Angiostrongylus vasorum through transcriptomics--prospects for new interventions.

    PubMed

    Ansell, Brendan R E; Schnyder, Manuela; Deplazes, Peter; Korhonen, Pasi K; Young, Neil D; Hall, Ross S; Mangiola, Stefano; Boag, Peter R; Hofmann, Andreas; Sternberg, Paul W; Jex, Aaron R; Gasser, Robin B

    2013-12-01

    Angiostrongylus vasorum is a metastrongyloid nematode of dogs and other canids of major clinical importance in many countries. In order to gain first insights into the molecular biology of this worm, we conducted the first large-scale exploration of its transcriptome, and predicted essential molecules linked to metabolic and biological processes as well as host immune responses. We also predicted and prioritized drug targets and drug candidates. Following Illumina sequencing (RNA-seq), 52.3 million sequence reads representing adult A. vasorum were assembled and annotated. The assembly yielded 20,033 contigs, which encoded proteins with 11,505 homologues in Caenorhabditis elegans, and additional 2252 homologues in various other parasitic helminths for which curated data sets were publicly available. Functional annotation was achieved for 11,752 (58.6%) proteins predicted for A. vasorum, including peptidases (4.5%) and peptidase inhibitors (1.6%), protein kinases (1.7%), G protein-coupled receptors (GPCRs) (1.5%) and phosphatases (1.2%). Contigs encoding excretory/secretory and immuno-modulatory proteins represented some of the most highly transcribed molecules, and encoded enzymes that digest haemoglobin were conserved between A. vasorum and other blood-feeding nematodes. Using an essentiality-based approach, drug targets, including neurotransmitter receptors, an important chemosensory ion channel and cysteine proteinase-3 were predicted in A. vasorum, as were associated small molecular inhibitors/activators. Future transcriptomic analyses of all developmental stages of A. vasorum should facilitate deep explorations of the molecular biology of this important parasitic nematode and support the sequencing of its genome. These advances will provide a foundation for exploring immuno-molecular aspects of angiostrongylosis and have the potential to underpin the discovery of new methods of intervention.

  18. [Influencing factors on culture of medicinal plants adventitious roots].

    PubMed

    Yin, Shuang-Shuang; Gao, Wen-Yuan; Wang, Juan; Liu, Hui; Zuo, Bei-Mei

    2012-12-01

    With the modernization of traditional Chinese medicine, medicinal plants resources cannot meet the request of Chinese medicine industry. Medicinal plants adventitious roots culture in a large scale is an important way to achieve Chinese medicine industrialization. However, how to establish good adventitious roots culture system is its key, such as plant hormones, explant, sucrose, innoculum and salt strength.

  19. [Influencing factors on culture of medicinal plants adventitious roots].

    PubMed

    Yin, Shuang-Shuang; Gao, Wen-Yuan; Wang, Juan; Liu, Hui; Zuo, Bei-Mei

    2012-12-01

    With the modernization of traditional Chinese medicine, medicinal plants resources cannot meet the request of Chinese medicine industry. Medicinal plants adventitious roots culture in a large scale is an important way to achieve Chinese medicine industrialization. However, how to establish good adventitious roots culture system is its key, such as plant hormones, explant, sucrose, innoculum and salt strength. PMID:23627161

  20. Comparison of faecal techniques including FLOTAC for copromicroscopic detection of first stage larvae of Angiostrongylus vasorum.

    PubMed

    Schnyder, Manuela; Maurelli, Maria P; Morgoglione, Maria E; Kohler, Lucia; Deplazes, Peter; Torgerson, Paul; Cringoli, Giuseppe; Rinaldi, Laura

    2011-07-01

    Angiostrongylus vasorum is a metastrongylid nematode that resides in the pulmonary arteries and the right heart chambers. In dogs, infection results in respiratory, bleeding and neurological disorders and further clinical signs. In the present study, FLOTAC was evaluated for the detection of first-stage larvae (L1) of A. vasorum in canine faecal samples. This technique is based on the counting of parasitic stages (eggs, larvae, oocysts and cysts) in chambers after spinning of faecal samples onto a surface. In a first step, nine flotation solutions were evaluated using faeces of two experimentally infected dogs. Zinc sulphate (specific gravity (s.g.) 1.2) and zinc sulphate plus potassium iodomercurate (s.g. 1.45) gave good results. However, with the latter technique, the larvae were slightly deformed. Subsequently, FLOTAC, using zinc sulphate, was compared through a randomisation technique with McMaster, flotation in tube and Baermann-Wetzel technique. The mean larvae per gramme (LPG) obtained by the FLOTAC for both dogs was significantly higher (P < 0.05) than those obtained by the other three techniques (the means of the other techniques all lie below the 95% CI of the mean LPG of the FLOTAC technique). In addition, the FLOTAC results were consistent across replicates with only Poisson (or random) variation between individual replicates. The other techniques appear to be less consistent with evidence of extra-Poisson variation in at least one of the two dogs across the replicates within each technique. The FLOTAC technique may contribute to an improvement of the ability to diagnose canine lungworm infections and represent a valuable alternative for larval counting of A. vasorum in faecal samples, especially following transport or storage where there may be limited larvae viability, and larval migration techniques cannot be used.

  1. Molecular cloning of vasa gene and the effects of LHRH-A on its expression in blue tilapia Oreochromis aureus.

    PubMed

    Xiao, Jun; Luo, Yongju; Chen, Libing; Yang, Li; Huang, Yulin; Guo, Zhongbao; Guo, Enyan; Tang, Zhanyang; Zhang, Ming; Gan, Xi

    2013-08-01

    The full length of vasa cDNA in blue tilapia Oreochromis aureus was cloned and sequenced using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Nucleotide sequence analysis revealed that the cDNA contained 2,143 bp and was consisted of a 48-bp 5' untranslated terminal region (5'-UTR), a 157-bp 3' untranslated terminal region (3'-UTR) and a 1,938-bp open reading frame (ORF) which encoded 645 amino acids. Homological protein analysis showed that vasa in O. aureus was highly conserved with Nile tilapia Oreochromis niloticus. Tissue distribution expression analysis indicated that vasa was specifically expressed in the gonads. Using in situ hybridization, we found that vasa was expressed in spermatogonia and spermatocytes rather than spermatids and sperm. In order to examine the influence of luteinizing hormone releasing hormone analog (LHRH-A) on vasa, the in vivo injections were performed different concentrations of LHRH-A. Our results showed that LHRH-A induced meiosis and down-regulated vasa mRNA expression. In summary, our results showed that vasa was specifically expressed in gonads and LHRH-A inhibited vasa expression in the testis. Our results also suggested that LHRH-A could regulate vasa gene expression in O. aureus testis. PMID:23224831

  2. Increased prevalence and geographic spread of the cardiopulmonary nematode Angiostrongylus vasorum in fox populations in Great Britain.

    PubMed

    Taylor, C S; Garcia Gato, R; Learmount, J; Aziz, N A; Montgomery, C; Rose, H; Coulthwaite, C L; McGarry, J W; Forman, D W; Allen, S; Wall, R; Morgan, E R

    2015-08-01

    The nematode Angiostrongylus vasorum is becoming more widely recorded globally, and is of increasing concern as a cause of disease in dogs. Apparent geographic spread is difficult to confirm due to a lack of standardized disease recording systems, increasing awareness among veterinary clinicians, and recent improvements in diagnostic technologies. This study examines the hypothesis that A. vasorum has spread in recent years by repeating the methods of a previous survey of the fox population. The hearts and lungs of 442 foxes from across Great Britain were collected and examined by dissection and flushing of the pulmonary circulation and microscopic inspection of tracheal scrapes. Sampling and parasite extraction methods were identical to an earlier survey in 2005 to ensure comparability. Prevalence of A. vasorum was 18·3% (exact binomial confidence bounds 14·9-22·3), compared with 7·3% previously (5·3-9·9, n = 546), and had increased significantly in most regions, e.g. 7·4% in the Northern UK (previously zero) and 50·8% in the south-east (previously 23·2%). Other nematodes identified were Crenosoma vulpis (prevalence 10·8%, CI 8·1-14·2) and Eucoleus aerophilus (31·6%, CI 27·3-36·2). These data support the proposal that A. vasorum has increased in prevalence and has spread geographically in Great Britain.

  3. Angiostrongylus vasorum in Great Britain: a nationwide postal questionnaire survey of veterinary practices.

    PubMed

    Kirk, L; Limon, G; Guitian, F J; Hermosilla, C; Fox, M T

    2014-08-01

    The lungworm, Angiostrongylus vasorum, was first reported in indigenous dogs in southwestern England in 1980 and has since been recorded in Wales, southeastern England and, more recently, in the West Midlands, northern England and Scotland. The nationwide distribution of the parasite was evaluated using a postal questionnaire sent to 3950 small animal practices during 2009. Information was sought on the location of each practice, awareness of the parasite locally, number of cases diagnosed over the past year and whether diagnosis was based on clinical signs alone or supported by additional tests. 1419 practices returned a usable response, the majority being located in a city/town. Nearly one-third of responding practices were aware of the parasite locally, 20.7 per cent had seen at least one confirmed case and 0.3 per cent >20 confirmed cases over the past year. The most widely used tests were faecal examination and any type of imaging. Existing clusters of infection were detected in southeastern England and south Wales; infection was also found to be widespread in central England, though patchy in northern England and Scotland. Using distribution of clinical cases as an indicator of parasite distribution, this study confirmed that A. vasorum has spread beyond traditional UK endemic foci.

  4. Spatial, demographic and clinical patterns of Angiostrongylus vasorum infection in the dog population of Southern England.

    PubMed

    Blehaut, T R W; Hardstaff, J L; Chapman, P S; Pfeiffer, D U; Boag, A K; Guitian, F J

    2014-08-01

    A retrospective study was carried out to provide updated knowledge of the spatial pattern of Angiostrongylus vasorum infection in Southern England and to investigate associations between selected host characteristics (age, breed, sex), risk of infection and clinical presentation (cardiorespiratory signs v haemorrhagic diathesis). One hundred and forty-one cases diagnosed between April 1999 and July 2012 were compared with a control population of dogs referred to the same hospital. A significant association was found between haemorrhagic diathesis and breed but not for other host characteristics and clinical presentations. Younger dogs and certain breeds of dog (Jack Russell terriers, Cocker Spaniels, Springer Spaniels, Cavalier King Charles spaniels and Staffordshire Bull Terriers) had significantly higher odds of angiostrongylosis than other breeds in the study. A significant cluster of cases was found in Southern England. Animals presenting with cardiorespiratory signs or haemorrhagic diathesis in Southern England, especially if they are young or of a breed associated with angiostrongylosis, should be given special consideration with regards to possible A. vasorum infestation. Our results should be interpreted bearing in mind that they are based on the retrospective exploration of dogs seen at a referral centre.

  5. Identification and sexually dimorphic expression of vasa isoforms in Dabry's sturgeon (Acipenser dabryanus), and functional analysis of vasa 3'-untranslated region.

    PubMed

    Ye, Huan; Yue, Hua-Mei; Yang, Xiao-Ge; Li, Chuang-Ju; Wei, Qi-Wei

    2016-10-01

    Germ cells are set aside from somatic cells early in embryogenesis, and are responsible for transmitting genetic information through generations. Vasa is a highly conserved germ cell marker across animal phyla, and widely used to label primordial germ cells. Dabry's sturgeon is a rare and endangered species distributed solely in the Yangtze River basin. Here, seven vasa isoforms, named Advasa1-7, were isolated and characterized in Dabry's sturgeon. RT-PCR and western blot analyses revealed that vasa mRNA and protein were mainly restricted to the testis and ovary, but exhibited sexually dimorphic expression. Cellular and subcellular localization uncovered that Advasa mRNA and protein displayed mitotic and meiotic expression in females, and mainly showed mitotic expression in males; surprisingly, they exhibited both cytoplasmic and nuclear expression in the ovarian germ cells, while showing exclusively cytoplasmic expression in the testicular germ cells. By microinjecting chimeric RNA consisting of the red fluorescent protein coding region and the Advasa 3'-untranslated region into embryos of Dabry's sturgeon, zebrafish and medaka, we demonstrated that it had the ability to visualize primordial germ cells (PGCs) in Dabry's sturgeon and zebrafish but not in medaka. It seemed that the machinery of vasa 3'UTR RNA localization was conserved between Dabry's sturgeon and ostariophysan, while possibly changed during the divergence of euteleosts and ostariophysan. Finally, Dabry's sturgeon PGCs moved on the yolk ball, and migrated toward the genital ridge via mesenchyme. Taken together, these results provide new information for vasa expression pattern and function, and lay a foundation for PGC cryopreservation and conservation of Dabry's sturgeon.

  6. Combined Serological Detection of Circulating Angiostrongylus vasorum Antigen and Parasite-specific Antibodies in Dogs from Hungary.

    PubMed

    Schnyder, Manuela; Schaper, Roland; Lukács, Zoltán; Hornok, Sándor; Farkas, Róbert

    2015-08-01

    The occurrence of the nematode Angiostrongylus vasorum, also known as the French heartworm, is increasingly being reported from various European countries. The adults of this parasite species live in the pulmonary arteries and right cardiac ventricle of wild canids and domestic dogs. Larval stages and eggs in the lungs induce inflammatory verminous pneumonia, causing severe respiratory disease in dogs. Furthermore, haematological and neurological signs and even death may occur. In Hungary, A. vasorum has been identified in red foxes, golden jackals and in two dogs and some slugs. In this first large-scale survey, 1247 sera from pet dogs were collected and tested by an ELISA for the detection of circulating antigen of A. vasorum and by a separate ELISA to detect specific antibodies against the parasite. A total of 1.36% (n = 17, 95 % confidence intervals, CI: 0.80 - 2.17 %) of the animals were positive in both ELISAs, while 1.76 % (n = 22, CI: 1.11 - 2.66 %) of the tested dogs were antigen-positive only and 2.73 % (n = 34, CI: 1.90 - 3.79 %) were positive for specific antibodies only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly the whole sampled areas of the country. A considerable number of cases was observed in Budapest and also in the southern part of the country bordering Croatia, while in the most eastern part bordering Ukraine no positive samples were detected. These results confirm the endemic occurrence of A. vasorum in dogs originating from different parts of Hungary and the significant advantages of A. vasorum serology in epidemiological studies. PMID:26152415

  7. Combined Serological Detection of Circulating Angiostrongylus vasorum Antigen and Parasite-specific Antibodies in Dogs from Hungary.

    PubMed

    Schnyder, Manuela; Schaper, Roland; Lukács, Zoltán; Hornok, Sándor; Farkas, Róbert

    2015-08-01

    The occurrence of the nematode Angiostrongylus vasorum, also known as the French heartworm, is increasingly being reported from various European countries. The adults of this parasite species live in the pulmonary arteries and right cardiac ventricle of wild canids and domestic dogs. Larval stages and eggs in the lungs induce inflammatory verminous pneumonia, causing severe respiratory disease in dogs. Furthermore, haematological and neurological signs and even death may occur. In Hungary, A. vasorum has been identified in red foxes, golden jackals and in two dogs and some slugs. In this first large-scale survey, 1247 sera from pet dogs were collected and tested by an ELISA for the detection of circulating antigen of A. vasorum and by a separate ELISA to detect specific antibodies against the parasite. A total of 1.36% (n = 17, 95 % confidence intervals, CI: 0.80 - 2.17 %) of the animals were positive in both ELISAs, while 1.76 % (n = 22, CI: 1.11 - 2.66 %) of the tested dogs were antigen-positive only and 2.73 % (n = 34, CI: 1.90 - 3.79 %) were positive for specific antibodies only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly the whole sampled areas of the country. A considerable number of cases was observed in Budapest and also in the southern part of the country bordering Croatia, while in the most eastern part bordering Ukraine no positive samples were detected. These results confirm the endemic occurrence of A. vasorum in dogs originating from different parts of Hungary and the significant advantages of A. vasorum serology in epidemiological studies.

  8. Adventitial inflammation and its interaction with intimal atherosclerotic lesions

    PubMed Central

    Akhavanpoor, Mohammadreza; Wangler, Susanne; Gleissner, Christian A.; Korosoglou, Grigorios; Katus, Hugo A.; Erbel, Christian

    2014-01-01

    The presence of adventitial inflammation in correlation with atherosclerotic lesions has been recognized for decades. In the last years, several studies have investigated the relevance and impact of adventitial inflammation on atherogenesis. In the abdominal aorta of elderly Apoe−/− mice, adventitial inflammatory structures were characterized as organized ectopic lymphoid tissue, and therefore termed adventitial tertiary lymphoid organs (ATLOs). These ATLOs possess similarities in development, structure and function to secondary lymphoid organs. A crosstalk between intimal atherosclerotic lesions and ATLOs has been suggested, and several studies could demonstrate a potential role for medial vascular smooth muscle cells in this process. We here review the development, phenotypic characteristics, and function of ATLOs in atherosclerosis. Furthermore, we discuss the possible role of medial vascular smooth muscle cells and their interaction between plaque and ATLOs. PMID:25152736

  9. The characteristics of vasa gene from Japanese sea bass ( Lateolabrax japonicas) and its response to the external hormones

    NASA Astrophysics Data System (ADS)

    Chi, Meili; Wen, Haishen; Ni, Meng; Qian, Kun; Zhang, Pei; Chai, Senhao

    2015-08-01

    The RNA helicase Vasa is an important regulator of primordial germ cell development. Its function in mature fish, especially the hormone-related differences in maturing male fish has seldom been documented. In this study, a full length cDNA sequence of the vasa gene was cloned from Japanese sea bass, Lateolabrax japonicas, and it was named jsb-vasa. Homology analysis showed that jsb-vasa was closely related to its teleost homologs. The spatial distribution of jsb-vasa indicated that it was only highly expressed in testis, showing its germ cell-specific expression pattern. During the testicular development cycle, jsb-vasa was highly expressed during early period of spermatogenesis, and reduced when spermatogenesis advanced. In addition, the jsb-vasa gene expression was significantly inhibited at 6 h, 12 h and 24 h after injecting hCG (human chorionic gonadotropin) and GnRHa (Gonadotropin-releasing hormone analogue), indicating that jsb-vasa gene may play an important role in spermatogenesis of Japanese sea bass, and be under the regulation of external sex hormones.

  10. An unregulated regulator: Vasa expression in the development of somatic cells and in tumorigenesis.

    PubMed

    Poon, Jessica; Wessel, Gary M; Yajima, Mamiko

    2016-07-01

    Growing evidence in diverse organisms shows that genes originally thought to function uniquely in the germ line may also function in somatic cells, and in some cases even contribute to tumorigenesis. Here we review the somatic functions of Vasa, one of the most conserved "germ line" factors among metazoans. Vasa expression in somatic cells is tightly regulated and often transient during normal development, and appears to play essential roles in regulation of embryonic cells and regenerative tissues. Its dysregulation, however, is believed to be an important element of tumorigenic cell regulation. In this perspectives paper, we propose how some conserved functions of Vasa may be selected for somatic cell regulation, including its potential impact on efficient and localized translational activities and in some cases on cellular malfunctioning and tumorigenesis. PMID:27179696

  11. vasa and piwi are required for mitotic integrity in early embryogenesis in the spider Parasteatoda tepidariorum.

    PubMed

    Schwager, Evelyn E; Meng, Yue; Extavour, Cassandra G

    2015-06-15

    Studies in vertebrate and invertebrate model organisms on the molecular basis of primordial germ cell (PGC) specification have revealed that metazoans can specify their germ line either early in development by maternally transmitted cytoplasmic factors (inheritance), or later in development by signaling factors from neighboring tissues (induction). Regardless of the mode of PGC specification, once animal germ cells are specified, they invariably express a number of highly conserved genes. These include vasa and piwi, which can play essential roles in any or all of PGC specification, development, or gametogenesis. Although the arthropods are the most speciose animal phylum, to date there have been no functional studies of conserved germ line genes in species of the most basally branching arthropod clade, the chelicerates (which includes spiders, scorpions, and horseshoe crabs). Here we present the first such study by using molecular and functional tools to examine germ line development and the roles of vasa and piwi orthologues in the common house spider Parasteatoda (formerly Achaearanea) tepidariorum. We use transcript and protein expression patterns of Pt-vasa and Pt-piwi to show that primordial germ cells (PGCs) in the spider arise during late embryogenesis. Neither Pt-vasa nor Pt-piwi gene products are localized asymmetrically to any embryonic region before PGCs emerge as paired segmental clusters in opisthosomal segments 2-6 at late germ band stages. RNA interference studies reveal that both genes are required maternally for egg laying, mitotic progression in early embryos, and embryonic survival. Our results add to the growing body of evidence that vasa and piwi can play important roles in somatic development, and provide evidence for a previously hypothesized conserved role for vasa in cell cycle progression. PMID:25257304

  12. vasa and piwi are required for mitotic integrity in early embryogenesis in the spider Parasteatoda tepidariorum.

    PubMed

    Schwager, Evelyn E; Meng, Yue; Extavour, Cassandra G

    2015-06-15

    Studies in vertebrate and invertebrate model organisms on the molecular basis of primordial germ cell (PGC) specification have revealed that metazoans can specify their germ line either early in development by maternally transmitted cytoplasmic factors (inheritance), or later in development by signaling factors from neighboring tissues (induction). Regardless of the mode of PGC specification, once animal germ cells are specified, they invariably express a number of highly conserved genes. These include vasa and piwi, which can play essential roles in any or all of PGC specification, development, or gametogenesis. Although the arthropods are the most speciose animal phylum, to date there have been no functional studies of conserved germ line genes in species of the most basally branching arthropod clade, the chelicerates (which includes spiders, scorpions, and horseshoe crabs). Here we present the first such study by using molecular and functional tools to examine germ line development and the roles of vasa and piwi orthologues in the common house spider Parasteatoda (formerly Achaearanea) tepidariorum. We use transcript and protein expression patterns of Pt-vasa and Pt-piwi to show that primordial germ cells (PGCs) in the spider arise during late embryogenesis. Neither Pt-vasa nor Pt-piwi gene products are localized asymmetrically to any embryonic region before PGCs emerge as paired segmental clusters in opisthosomal segments 2-6 at late germ band stages. RNA interference studies reveal that both genes are required maternally for egg laying, mitotic progression in early embryos, and embryonic survival. Our results add to the growing body of evidence that vasa and piwi can play important roles in somatic development, and provide evidence for a previously hypothesized conserved role for vasa in cell cycle progression.

  13. Seroepidemiological survey and spatial analysis of the occurrence of Angiostrongylus vasorum in Swiss dogs in relation to biogeographic aspects.

    PubMed

    Lurati, L; Deplazes, P; Hegglin, D; Schnyder, M

    2015-09-15

    Angiostrongylus vasorum is a metastrongyloid nematode living in the pulmonary arteries and in the right heart causing potentially fatal respiratory distress, coagulopathies and a wide range of other clinical signs in dogs. The aim of this study was to investigate the seroprevalence and distribution of A. vasorum in Swiss dogs and to identify correlations with biogeographic aspects. A total of 6136 dog sera from all over the country submitted by veterinarians for haematological or chemical analyses were examined for the presence of circulating A. vasorum parasite antigen and specific antibodies against A. vasorum by ELISA. The combined seroprevalence for both specific antibodies and antigen was 0.96% (95% confidence intervals: 0.7-1.2%), while the overall antibody prevalence was 3.08% (CI: 2.7-3.5%) and the antigen prevalence 2.17% (CI: 1.8-2.6%). The highest prevalence for dogs identified as positive in both ELISAs was detected in Western Switzerland (around Geneva, 2.21%, CI: 0.7-5.1%), representing a new endemic area. Known endemic regions in Southern Switzerland (Ticino, 2.17%, CI: 1.0-4.0%) and in the High Rhine area (northern Switzerland, 1.11%, CI: 0.4-2.4%) were confirmed. Spatial analysis identified a cluster with a radius of approximately 30 km in Sisseln, located in this latter region at the German border, for antibody positive dogs, which interestingly corresponded to the location of historical cases of canine angiostrongylosis diagnosed 12-16 years previously. In total 96.6% (57/59) of the antigen- and antibody positive samples originated from areas with a mean temperature warmer than -2 °C in January. Correspondingly, most of the samples (53/59, 89.8%) positive in both ELISAs originated from areas below 700 meters above sea level (m asl), suggesting the altitude being a limiting factor for A. vasorum transmission in Switzerland. This study confirms previously known endemic areas for A. vasorum in Switzerland, and additionally identifies positive dogs

  14. Emerging from the clouds: Vasa helicase sheds light on piRNA amplification.

    PubMed

    Claycomb, Julie M

    2014-06-23

    Reporting in a recent study in Cell, Xiol and colleagues (2014) identify the conserved DEAD box helicase Vasa as a platform for secondary Piwi-interacting RNA (piRNA) biogenesis in the insect nuage. This study represents a substantial breakthrough in understanding the mechanism and location of piRNA amplification by the "ping-pong" cycle.

  15. Predatory activity of the fungi Duddingtonia flagrans, Monacrosporium thaumasium, Monacrosporium sinense and Arthrobotrys robusta on Angiostrongylus vasorum first-stage larvae.

    PubMed

    Braga, F R; Carvalho, R O; Araujo, J M; Silva, A R; Araújo, J V; Lima, W S; Tavela, A O; Ferreira, S R

    2009-12-01

    Angiostrongylus vasorum is a nematode that parasitizes domestic dogs and wild canids. We compared the predatory capacity of isolates from the predatory fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on first-stage larvae (L1) of A. vasorum under laboratory conditions. L1 A. vasorum were plated on 2% water-agar (WA) Petri dishes marked into 4 mm diameter fields with the four grown isolates and a control without fungus. Plates of treated groups contained each 1000 L1 A. vasorum and 1000 conidia of the fungal isolates AC001, NF34, SF53 and I31 on 2% WA. Plates of the control group (without fungus) contained only 1000 L1 A. vasorum on 2% WA. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for 7 days. Nematophagous fungi were not observed in the control group during the experiment. There was no variation in the predatory capacity among the tested fungal isolates (P>0.05) during the 7 days of the experiment. There was a significant reduction (P < 0.05) of 80.3%, 74.5%, 74.2% and 71.8% in the means of A. vasorum L1 recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively, compared to the control without fungi. In this study, the four isolates of predatory fungi were efficient in the in vitro capture and destruction of A. vasorum L1, confirming previous work on the efficiency of nematophagous fungi in the control of nematode parasites of dogs and as a possible alternative method of biological control. PMID:19216825

  16. Autologous adventitial overlay method reinforces anastomoses in aortic surgery.

    PubMed

    Minato, Naoki; Okada, Takayuki; Sumida, Tomohiko; Watanabe, Kenichi; Maruyama, Takahiro; Kusunose, Takashi

    2014-05-01

    In this study, we present an inexpensive and effective method for providing a secure and hemostatic anastomosis using autologous adventitia obtained from a dissected or aneurysmal wall. The resected aortic wall is separated between the adventitia and media, and a soft, 2 × 10-cm adventitial strip is overlaid to cover the anastomotic margin. A graft is sutured to the aortic stump. This autologous adventitial overlay method can inexpensively and strongly reinforce the anastomosis during aortic surgery for dissection or aneurysm and will contribute to anastomotic hemostasis and long-term stability.

  17. Molecular characterization and expression of buffalo (Bubalus bubalis) DEAD-box family VASA gene and mRNA transcript variants isolated from testis tissue.

    PubMed

    Kaushik, Ramakant; Singh, Karn Pratap; Bahuguna, Vivek; Rameshbabu, K; Singh, Manoj Kumar; Manik, Radhey Shyam; Palta, Prabhat; Singla, Suresh Kumar; Chauhan, Manmohan Singh

    2015-11-01

    VASA is a member of the DEAD-box protein family that plays an indispensable role in mammalian spermatogenesis, particularly during meiosis. In the present study, we isolated, sequenced, and characterized VASA gene in buffalo testis. Here, we demonstrated that VASA mRNA is expressed as multiple isoforms and uses four alternative transcriptional start sites (TSSs) and four different polyadenylation sites. The TSSs identified by 5'-RNA ligase-mediated rapid amplification of cDNA ends (RLM-5'-RACE) were positioned at 48, 53, 85, and 88 nucleotides upstream relative to the translation initiation codon. 3'-RACE experiment revealed the presence of tandem polyadenylation signals, which lead to the expression of at least four different 3'-untranslated regions (209, 233, 239 and 605 nucleotides). The full-length coding region of VASA was 2190 bp, which encodes a 729 amino acid (aa) protein containing nine consensus regions of the DEAD box protein family. VASA variants are highly expressed in testis of adult buffalo. We found five variants, one full length VASA (729 aa) and four splice variants VASA 2, 4, 5, 6 (683, 685, 679, 703 aa). The expression level of VASA 1 was significantly higher than rest of all (P < 0.05) except VASA 6. The relative ratio for VASA 1:2:4:5:6 was 100:1.0:1.6:0.9:48.

  18. [Natural infection with Angiostrongylus vasorum: characterisation of 3 dogs with pulmonary hypertension].

    PubMed

    Glaus, T; Schnyder, M; Dennler, M; Tschuor, F; Wenger, M; Sieber-Ruckstuhl, N

    2010-07-01

    Pulmonary hypertension (PH), together with its accompanying clinical signs and underlying causes, e.g. pulmonary thrombosis, are more and more recognized as an important clinical entity also in dogs. This article characterizes the clinical picture of 3 dogs with PH caused by natural infection with Angiostrongylus vasorum. All 3 dogs were of small breeds ( < 10 kg), the age at the time of diagnosis was 1, 2 and 11 years. Clinically, dyspnea and exercise intolerance were the predominating signs, 2 dogs developed hemoptysis, 1 dog developed right sided congestive heart failure. Severe arterial hypoxemia (PaO2 41 - 53 mmHg) reflected the severity of pulmonary parenchymal and vascular damage. Severe hyperglobulinemia (59 und 88 g/l) in two dogs implicated a long lasting infection. Anthelmintic treatment in 2 dogs resulted in quick clinical, radiographic and echocardiographic normalization. PH is the consequence of multiple causes and pathomechanisms, and the recognition of PH is primarily of differential diagnostic relevance. Prognosis and therapy in cases with PH mainly depend on the underlying cause, rather than on the PH and on its degree.

  19. Velamentous insertion of umbilical cord with vasa praevia: case series and literature review

    PubMed Central

    Bohîlțea, RE; Cîrstoiu, MM; Ciuvica, AI; Munteanu, O; Bodean, O; Voicu, D; Ionescu, CA

    2016-01-01

    A velamentous umbilical cord is characterized by membranous umbilical vessels at the placental insertion site that are prone to compression and rupture, especially when they are located in the membranes covering the cervical os (vasa praevia). The velamentous insertion of the umbilical cord, with a reported incidence of 1% in singleton pregnancies and 15% in monochorionic twin gestations, has been associated with obstetric complications: fetal growth restriction, prematurity, congenital anomalies, low Apgar scores, fetal bleeding with acute fetal distress and placental retention. The pathogenesis is unknown, but the trophotropism theory is the most common and supported by the association of velamentous cord insertion and placenta praevia. The prevalence of vasa praevia is of approximately 1/ 2500 deliveries; the risk factors include the use of assisted reproductive technologies, low-lying placenta or placenta praevia, bilobed or succenturiate lobe placenta and multiple gestation. The diagnosis is rarely established before delivery and consequently the fetal mortality is extremely high. We report two cases of velamentous marginal umbilical cord insertion associated with vasa praevia (type 1 vasa praevia) and placenta praevia diagnosed during a routine mid-trimester fetal 2D ultrasound scan, color and power Doppler transvaginal ultrasound cervical assessment. The ultrasound examination revealed one umbilical vessel crossing the internal os of the cervix entering the placental margin and connecting to the subchorionic vasculature, remaining immobile when the uterus was shaken, the color Doppler imaging enhancing the identification of the vessel. The patients were admitted to the hospital in the third trimester and deliveries were planed and successfully performed at 38 weeks gestation, being confirmed by a macroscopic examination ultrasound diagnostic. PMID:27453740

  20. Transport of sodium and urea in outer medullary descending vasa recta.

    PubMed Central

    Pallone, T L; Work, J; Myers, R L; Jamison, R L

    1994-01-01

    We dissected and perfused outer medullary vasa recta (OMVR) from vascular bundles in the rat. Permeabilities of sodium (PNa) and urea (Pu) were simultaneously determined from the lumen-to-bath efflux of 22Na and [14C]urea. PNa and Pu were also measured by in vivo microperfusion of descending (DVR) and ascending vasa recta (AVR) at the papillary tip of Munich-Wistar rats. In some OMVR PNa was indistinguishable from zero. The mean +/- SE of PNa (x 10(-5), cm/s) in OMVR was 76 +/- 9. Pu in OMVR was always very high (x 10(-5), cm/s), 360 +/- 14. There was no correlation between OMVR PNa and Pu. Inner medullary AVR and DVR had PNa of 115 +/- 10 and 75 +/- 10, respectively, and Pu of 121 +/- 10 and 76 +/- 11, respectively. PNa and Pu in papillary vasa recta were always nearly identical and highly correlated. Transport of [14C] urea in OMVR was reversibly inhibited by addition of unlabeled urea or phloretin to the bath and lumen, providing evidence for carrier-mediated transport. These data suggest that sodium and urea might traverse the wall of inner medullary vasa recta by a paracellular pathway while urea also crosses by a transcellular route in OMVR. Electron microscopic examination of seven in vitro perfused OMVR revealed no fenestrations and exposure of these vessels to 10 microM calcium ionophore A23187 or 1 nM angiotensin II resulted in reversible contraction, suggesting that in vitro perfused OMVR are DVR only. Images PMID:8282790

  1. Massively parallel sequencing, a new method for detecting adventitious agents.

    PubMed

    Onions, David; Kolman, John

    2010-05-01

    There has been an upsurge of interest in developing new veterinary and human vaccines and, in turn, this has involved the development of new mammalian and insect cell substrates. Excluding adventitious agents from these cells can be problematic, particularly for cells derived from species with limited virological investigation. Massively parallel sequencing is a powerful new method for the identification of viruses and other adventitious agents, without prior knowledge of the nature of the agent. We have developed methods using random priming to detect viruses in the supernatants from cell substrates or in virus seed stocks. Using these methods we have recently discovered a new parvovirus in bovine serum. When applied to sequencing the transcriptome, massively parallel sequencing can reveal latent or silent infections. Enormous amounts of data are developed in this process usually between 100 and 400 Mbp. Consequently, sophisticated bioinformatic algorithms are required to analyse and verify virus targets.

  2. Improvement of adventitious root formation in flax using hydrogen peroxide.

    PubMed

    Takáč, Tomáš; Obert, Bohuš; Rolčík, Jakub; Šamaj, Jozef

    2016-09-25

    Flax (Linum usitatissimum L.) is an important crop for the production of oil and fiber. In vitro manipulations of flax are used for genetic improvement and breeding while improvements in adventitious root formation are important for biotechnological programs focused on regeneration and vegetative propagation of genetically valuable plant material. Additionally, flax hypocotyl segments possess outstanding morphogenetic capacity, thus providing a useful model for the investigation of flax developmental processes. Here, we investigated the crosstalk between hydrogen peroxide and auxin with respect to reprogramming flax hypocotyl cells for root morphogenetic development. Exogenous auxin induced the robust formation of adventitious roots from flax hypocotyl segments while the addition of hydrogen peroxide further enhanced this process. The levels of endogenous auxin (indole-3-acetic acid; IAA) were positively correlated with increased root formation in response to exogenous auxin (1-Naphthaleneacetic acid; NAA). Histochemical staining of the hypocotyl segments revealed that hydrogen peroxide and peroxidase, but not superoxide, were positively correlated with root formation. Measurements of antioxidant enzyme activities showed that endogenous levels of hydrogen peroxide were controlled by peroxidases during root formation from hypocotyl segments. In conclusion, hydrogen peroxide positively affected flax adventitious root formation by regulating the endogenous auxin levels. Consequently, this agent can be applied to increase flax regeneration capacity for biotechnological purposes such as improved plant rooting. PMID:26921706

  3. Transcript expression profiling for adventitious roots of Panax ginseng Meyer.

    PubMed

    Subramaniyam, Sathiyamoorthy; Mathiyalagan, Ramya; Natarajan, Sathishkumar; Kim, Yu-Jin; Jang, Moon-Gi; Park, Jun-Hyung; Yang, Deok Chun

    2014-08-01

    Panax ginseng Meyer is one of the major medicinal plants in oriental countries belonging to the Araliaceae family which are the primary source for ginsenosides. However, very few genes were characterized for ginsenoside pathway, due to the limited genome information. Through this study, we obtained a comprehensive transcriptome from adventitious roots, which were treated with methyl jasmonic acids for different time points (control, 2h, 6h, 12h, and 24h) and sequenced by RNA 454 pyrosequencing technology. Reference transcriptome 39,304,529 (0.04GB) was obtained from 5,724,987,880 bases (5.7GB) of 22 libraries by de novo assembly and 35,266 (58.5%) transcripts were annotated with biological schemas (GO and KEGG). The digital gene expression patterns were obtained from in vitro grown adventitious root sequences which mapped to reference, from that, 3813 (6.3%) unique transcripts were involved in ≥2 fold up and downregulations. Finally, candidates for ginsenoside pathway genes were predicted from observed expression patterns. Among them, 30 transcription factors, 20 cytochromes, and 11 glycosyl transferases were predicted as ginsenoside candidates. These data can remarkably expand the existing transcriptome resources of Panax, especially to predict existence of gene networks in P. ginseng. The entity of the data provides a valuable platform to reveal more on secondary metabolism and abiotic stresses from P. ginseng in vitro grown adventitious roots.

  4. Improvement of adventitious root formation in flax using hydrogen peroxide.

    PubMed

    Takáč, Tomáš; Obert, Bohuš; Rolčík, Jakub; Šamaj, Jozef

    2016-09-25

    Flax (Linum usitatissimum L.) is an important crop for the production of oil and fiber. In vitro manipulations of flax are used for genetic improvement and breeding while improvements in adventitious root formation are important for biotechnological programs focused on regeneration and vegetative propagation of genetically valuable plant material. Additionally, flax hypocotyl segments possess outstanding morphogenetic capacity, thus providing a useful model for the investigation of flax developmental processes. Here, we investigated the crosstalk between hydrogen peroxide and auxin with respect to reprogramming flax hypocotyl cells for root morphogenetic development. Exogenous auxin induced the robust formation of adventitious roots from flax hypocotyl segments while the addition of hydrogen peroxide further enhanced this process. The levels of endogenous auxin (indole-3-acetic acid; IAA) were positively correlated with increased root formation in response to exogenous auxin (1-Naphthaleneacetic acid; NAA). Histochemical staining of the hypocotyl segments revealed that hydrogen peroxide and peroxidase, but not superoxide, were positively correlated with root formation. Measurements of antioxidant enzyme activities showed that endogenous levels of hydrogen peroxide were controlled by peroxidases during root formation from hypocotyl segments. In conclusion, hydrogen peroxide positively affected flax adventitious root formation by regulating the endogenous auxin levels. Consequently, this agent can be applied to increase flax regeneration capacity for biotechnological purposes such as improved plant rooting.

  5. Adventitious roots, leaf abscission and nutrient status of flooded Gmelina and Tectona seedlings.

    PubMed

    Osundina, M A; Osonubi, O

    1989-12-01

    When flooded, seedlings of Gmelina arborea Roxb. produced more adventitious roots, had lower foliar Mn concentrations and lost fewer leaves than seedlings of Tectona grandis L.f. Severing the adventitious roots produced by flooded Gmelina seedlings increased leaf Mn concentration and leaf abscission and reduced whole-plant dry matter production. Flooded Gmelina cuttings, which do not produce adventitious roots, abscised few leaves until foliar concentrations of Mn and Fe had risen substantially above those of unflooded cuttings, at which time most leaves were shed. The results indicate that the development of adventitious roots in flooded seedlings of Gmelina suppressed uptake of Mn thereby minimizing leaf abscission. PMID:14972970

  6. Phytochemical deterrence of snowshoe hare browsing by adventitious shoots of four alaskan trees.

    PubMed

    Bryant, J P

    1981-08-21

    After snowshoe hares have severely browsed deciduous trees and shrubs, these woody plants produce adventitious shoots that are extremely unpalatable to them. The adventitious shoots of four common boreal forest trees contain significantly higher concentrations of terpene and phenolic resins than the mature-growth-form twigs of the same species. These resins are experimentally shown to be repellent to snowshoe hares and appear to explain the avoidance of adventitious shoots by hares. The production of adventitious shoots after intense hare browsing and the avoidance of these shoots by hares may play an important role in the 10-year hare cycle.

  7. Vascular autorescaling of fMRI (VasA fMRI) improves sensitivity of population studies: A pilot study

    PubMed Central

    Kazan, Samira M.; Mohammadi, Siawoosh; Callaghan, Martina F.; Flandin, Guillaume; Huber, Laurentius; Leech, Robert; Kennerley, Aneurin; Windischberger, Christian; Weiskopf, Nikolaus

    2016-01-01

    The blood oxygenation level-dependent (BOLD) signal is widely used for functional magnetic resonance imaging (fMRI) of brain function in health and disease. The statistical power of fMRI group studies is significantly hampered by high inter-subject variance due to differences in baseline vascular physiology. Several methods have been proposed to account for physiological vascularization differences between subjects and hence improve the sensitivity in group studies. However, these methods require the acquisition of additional reference scans (such as a full resting-state fMRI session or ASL-based calibrated BOLD). We present a vascular autorescaling (VasA) method, which does not require any additional reference scans. VasA is based on the observation that slow oscillations (< 0.1 Hz) in arterial blood CO2 levels occur naturally due to changes in respiration patterns. These oscillations yield fMRI signal changes whose amplitudes reflect the blood oxygenation levels and underlying local vascularization and vascular responsivity. VasA estimates proxies of the amplitude of these CO2-driven oscillations directly from the residuals of task-related fMRI data without the need for reference scans. The estimates are used to scale the amplitude of task-related fMRI responses, to account for vascular differences. The VasA maps compared well to cerebrovascular reactivity (CVR) maps and cerebral blood volume maps based on vascular space occupancy (VASO) measurements in four volunteers, speaking to the physiological vascular basis of VasA. VasA was validated in a wide variety of tasks in 138 volunteers. VasA increased t-scores by up to 30% in specific brain areas such as the visual cortex. The number of activated voxels was increased by up to 200% in brain areas such as the orbital frontal cortex while still controlling the nominal false-positive rate. VasA fMRI outperformed previously proposed rescaling approaches based on resting-state fMRI data and can be readily applied to

  8. VASA: Interactive Computational Steering of Large Asynchronous Simulation Pipelines for Societal Infrastructure.

    PubMed

    Ko, Sungahn; Zhao, Jieqiong; Xia, Jing; Afzal, Shehzad; Wang, Xiaoyu; Abram, Greg; Elmqvist, Niklas; Kne, Len; Van Riper, David; Gaither, Kelly; Kennedy, Shaun; Tolone, William; Ribarsky, William; Ebert, David S

    2014-12-01

    We present VASA, a visual analytics platform consisting of a desktop application, a component model, and a suite of distributed simulation components for modeling the impact of societal threats such as weather, food contamination, and traffic on critical infrastructure such as supply chains, road networks, and power grids. Each component encapsulates a high-fidelity simulation model that together form an asynchronous simulation pipeline: a system of systems of individual simulations with a common data and parameter exchange format. At the heart of VASA is the Workbench, a visual analytics application providing three distinct features: (1) low-fidelity approximations of the distributed simulation components using local simulation proxies to enable analysts to interactively configure a simulation run; (2) computational steering mechanisms to manage the execution of individual simulation components; and (3) spatiotemporal and interactive methods to explore the combined results of a simulation run. We showcase the utility of the platform using examples involving supply chains during a hurricane as well as food contamination in a fast food restaurant chain. PMID:26356899

  9. Vasa, PL10, and Piwi gene expression during caudal regeneration of the polychaete annelid Alitta virens.

    PubMed

    Kozin, Vitaly V; Kostyuchenko, Roman P

    2015-06-01

    Polychaetes are famous for their outstanding ability to regenerate lost body parts. Moreover, these worms possess a number of ancestral features in anatomy, development, and genetics, making them particularly suitable for comparative studies. Thus, fundamental as well as new undisclosed so far features of regenerative processes may be revealed, using polychaetes as a model. In the present work, we aimed to analyze the molecular basis of caudal regeneration in the nereid polychaete Alitta virens (formerly Nereis virens). We focused on homologues genes of RNA helicases Vasa and PL10 and ncRNA-binding proteins Piwi. These markers are suggested to play a significant role in maintenance of undifferentiated state of primordial germ cells and multipotent stem cells across invertebrates. In normal conditions, A. virens homologues of Vasa, PL10, and Piwi were differentially expressed in the subterminal growth zone and germline cells. Caudal amputation induced expression of studied genes de novo, which further accompanies all steps of regeneration. An early appearance of the transcripts in wound epithelium and internal blastemal cells suggests involvement of these genes in the well-known cell dedifferentiation events that assure polychaete regeneration. Provided interpretation of the gene expression dynamics implies the primary restoration of the pygidium and growth zone, which promotes following segment formation. Obtained results are valuable as a molecular fingerprint of the alterations occurring in regulatory state of locally regenerating tissues. PMID:25772273

  10. VASA: Interactive Computational Steering of Large Asynchronous Simulation Pipelines for Societal Infrastructure.

    PubMed

    Ko, Sungahn; Zhao, Jieqiong; Xia, Jing; Afzal, Shehzad; Wang, Xiaoyu; Abram, Greg; Elmqvist, Niklas; Kne, Len; Van Riper, David; Gaither, Kelly; Kennedy, Shaun; Tolone, William; Ribarsky, William; Ebert, David S

    2014-12-01

    We present VASA, a visual analytics platform consisting of a desktop application, a component model, and a suite of distributed simulation components for modeling the impact of societal threats such as weather, food contamination, and traffic on critical infrastructure such as supply chains, road networks, and power grids. Each component encapsulates a high-fidelity simulation model that together form an asynchronous simulation pipeline: a system of systems of individual simulations with a common data and parameter exchange format. At the heart of VASA is the Workbench, a visual analytics application providing three distinct features: (1) low-fidelity approximations of the distributed simulation components using local simulation proxies to enable analysts to interactively configure a simulation run; (2) computational steering mechanisms to manage the execution of individual simulation components; and (3) spatiotemporal and interactive methods to explore the combined results of a simulation run. We showcase the utility of the platform using examples involving supply chains during a hurricane as well as food contamination in a fast food restaurant chain.

  11. Hypocotyl adventitious root organogenesis differs from lateral root development

    PubMed Central

    Verstraeten, Inge; Schotte, Sébastien; Geelen, Danny

    2014-01-01

    Wound-induced adventitious root (AR) formation is a requirement for plant survival upon root damage inflicted by pathogen attack, but also during the regeneration of plant stem cuttings for clonal propagation of elite plant varieties. Yet, adventitious rooting also takes place without wounding. This happens for example in etiolated Arabidopsis thaliana hypocotyls, in which AR initiate upon de-etiolation or in tomato seedlings, in which AR initiate upon flooding or high water availability. In the hypocotyl AR originate from a cell layer reminiscent to the pericycle in the primary root (PR) and the initiated AR share histological and developmental characteristics with lateral roots (LRs). In contrast to the PR however, the hypocotyl is a determinate structure with an established final number of cells. This points to differences between the induction of hypocotyl AR and LR on the PR, as the latter grows indeterminately. The induction of AR on the hypocotyl takes place in environmental conditions that differ from those that control LR formation. Hence, AR formation depends on differentially regulated gene products. Similarly to AR induction in stem cuttings, the capacity to induce hypocotyl AR is genotype-dependent and the plant growth regulator auxin is a key regulator controlling the rooting response. The hormones cytokinins, ethylene, jasmonic acid, and strigolactones in general reduce the root-inducing capacity. The involvement of this many regulators indicates that a tight control and fine-tuning of the initiation and emergence of AR exists. Recently, several genetic factors, specific to hypocotyl adventitious rooting in A. thaliana, have been uncovered. These factors reveal a dedicated signaling network that drives AR formation in the Arabidopsis hypocotyl. Here we provide an overview of the environmental and genetic factors controlling hypocotyl-born AR and we summarize how AR formation and the regulating factors of this organogenesis are distinct from LR

  12. Cystic adventitial disease of the common femoral vein.

    PubMed

    Jayaraj, Arjun; Shalhub, Sherene; Deubner, Heike; Starnes, Benjamin W

    2011-05-01

    Cystic adventitial disease of blood vessels is a rare condition, more so when veins are involved. We report the case of a 36-year-old man who was referred to us after an intraoperative diagnosis of a left common femoral vein mass. This patient, who had a history of deep vein thrombosis and pulmonary embolism, had presented to an outside facility with recurrent left lower extremity pain and swelling. At our hospital, he underwent excision of the vein mass with interposition vein grafting using the left internal jugular vein. In this report, we discuss the presentation, diagnosis, treatment, and pathology of this rare condition. PMID:21549936

  13. Cystic Adventitial Disease of Popliteal Artery: Case Report.

    PubMed

    Meecham, Lewis; Wright, Anna; Atwal, Amarjit

    2016-03-01

    Cystic adventitial disease (CAD) is a rare cause of arterial stenosis. A total of 85% of cases are usually found in the popliteal artery. Men in their fourth and fifth decade are more commonly affected. If untreated the disease will progress eventually ending in occlusion of the affected vessel and limb-threatening ischemia. With this in mind an effective and long-lasting treatment is required, and here we present a case with CAD of the popliteal artery and discuss the merits of different treatment strategies.

  14. Identifying the germline in an equally cleaving mollusc: Vasa and Nanos expression during embryonic and larval development of the vetigastropod Haliotis asinina.

    PubMed

    Kranz, Alexandrea M; Tollenaere, Alina; Norris, Belinda J; Degnan, Bernard M; Degnan, Sandie M

    2010-06-15

    Members of the Vasa and Nanos gene families are important for the specification and development of the germline in diverse animals. Here, we determine spatial and temporal expression of Vasa and Nanos to investigate germline development in the vetigastropod Haliotis asinina. This is the first time these genes have been examined in an equally cleaving lophotrochozoan species. We find that HasVasa and HasNanos have largely overlapping, but not identical, expression patterns during embryonic and larval development, with both being maternally expressed and localized to the micromere cell lineages during cleavage. As embryonic development continues, HasVasa and HasNanos become progressively more enriched in the dorsal quadrant of the embryo. By the trochophore stage, both HasVasa and HasNanos are expressed in the putative mesodermal bands of the larva. This differs from the unequally cleaving gastropod Illyanasa obsoleta, in which IoVasa and IoNanos expression is detectable only in the early embryo and not during gastrulation and larval development. Our results suggest that the H. asinina germline arises from the 4d cell lineage and that primordial germ cells (PGCs) are not specified exclusively by maternally inherited determinants (preformation). As such, we infer that inductive signals (epigenesis) play an important role in specifying PGCs in H. asinina. We hypothesize that HasVasa is expressed in a population of undifferentiated multipotent cells, from which the PGCs are segregated later during development.

  15. In vitro regeneration of Salix nigra from adventitious shoots.

    PubMed

    Lyyra, Satu; Lima, Amparo; Merkle, Scott A

    2006-07-01

    Black willow (Salix nigra Marsh.) is the largest and only commercially important willow species in North America. It is a candidate for phytoremediation of polluted soils because it is fast-growing and thrives on floodplains throughout eastern USA. Our objective was to develop a protocol for the in vitro regeneration of black willow plants that could serve as target material for gene transformation. Unexpanded inflorescence explants were excised from dormant buds collected from three source trees and cultured on woody plant medium (WPM) supplemented with one of: (1) 0.1 mg l(-1) thidiazuron (TDZ); (2) 0.5 mg l(-1) 6-benzoaminopurine (BAP); or (3) 1 mg l(-1) BAP. All plant growth regulator (PGR) treatments induced direct adventitious bud formation from the genotypes. The percentage of explants producing buds ranged from 20 to 92%, depending on genotype and treatment. Although most of the TDZ-treated inflorescences produced buds, these buds failed to elongate into shoots. Buds on explants treated with BAP elongated into shoots that were easily rooted in vitro and further established in potting mix in high humidity. The PGR treatments significantly affected shoot regeneration frequency (P < 0.01). The highest shoot regeneration frequency (36%) was achieved with Genotype 3 cultured on 0.5 mg l(-1) BAP. Mean number of shoots per explant varied from one to five. The ability of black willow inflorescences to produce adventitious shoots makes them potential targets for Agrobacterium-mediated transformation with heavy-metal-resistant genes for phytoremediation.

  16. Expressed sequence tag analysis of functional genes associated with adventitious rooting in Liriodendron hybrids.

    PubMed

    Zhong, Y D; Sun, X Y; Liu, E Y; Li, Y Q; Gao, Z; Yu, F X

    2016-06-24

    Liriodendron hybrids (Liriodendron chinense x L. tulipifera) are important landscaping and afforestation hardwood trees. To date, little genomic research on adventitious rooting has been reported in these hybrids, as well as in the genus Liriodendron. In the present study, we used adventitious roots to construct the first cDNA library for Liriodendron hybrids. A total of 5176 expressed sequence tags (ESTs) were generated and clustered into 2921 unigenes. Among these unigenes, 2547 had significant homology to the non-redundant protein database representing a wide variety of putative functions. Homologs of these genes regulated many aspects of adventitious rooting, including those for auxin signal transduction and root hair development. Results of quantitative real-time polymerase chain reaction showed that AUX1, IRE, and FB1 were highly expressed in adventitious roots and the expression of AUX1, ARF1, NAC1, RHD1, and IRE increased during the development of adventitious roots. Additionally, 181 simple sequence repeats were identified from 166 ESTs and more than 91.16% of these were dinucleotide and trinucleotide repeats. To the best of our knowledge, the present study reports the identification of the genes associated with adventitious rooting in the genus Liriodendron for the first time and provides a valuable resource for future genomic studies. Expression analysis of selected genes could allow us to identify regulatory genes that may be essential for adventitious rooting.

  17. Adventitious Carbon on Primary Sample Containment Metal Surfaces

    NASA Technical Reports Server (NTRS)

    Calaway, M. J.; Fries, M. D.

    2015-01-01

    Future missions that return astromaterials with trace carbonaceous signatures will require strict protocols for reducing and controlling terrestrial carbon contamination. Adventitious carbon (AC) on primary sample containers and related hardware is an important source of that contamination. AC is a thin film layer or heterogeneously dispersed carbonaceous material that naturally accrues from the environment on the surface of atmospheric exposed metal parts. To test basic cleaning techniques for AC control, metal surfaces commonly used for flight hardware and curating astromaterials at JSC were cleaned using a basic cleaning protocol and characterized for AC residue. Two electropolished stainless steel 316L (SS- 316L) and two Al 6061 (Al-6061) test coupons (2.5 cm diameter by 0.3 cm thick) were subjected to precision cleaning in the JSC Genesis ISO class 4 cleanroom Precision Cleaning Laboratory. Afterwards, the samples were analyzed by X-ray photoelectron spectroscopy (XPS) and Raman spectroscopy.

  18. Mineral nutrition and adventitious rooting in microcuttings of Eucalyptus globulus.

    PubMed

    Schwambach, Joséli; Fadanelli, Cristina; Fett-Neto, Arthur G

    2005-04-01

    We characterized the adventitious rooting response of Eucalyptus globulus Labill. to various concentrations of calcium, nitrogen, phosphorus, iron, manganese, zinc, boron and copper. The parameters analyzed were percent rooting, root number, root length and mean rooting time. Root number and root length were significantly affected by mineral nutrition, whereas mean rooting time and rooting percentage seemed to be closely related to auxin availability. Root number was affected by calcium, nitrogen source and zinc, whereas root length was influenced by concentrations of phosphorus, iron and manganese, and by nitrogen source. Based on these results, we evaluated various combinations of several concentrations of these minerals in each rooting phase. Cuttings that were rooted in an optimized mineral nutrient medium and acclimatized to ex-vitro conditions for two months showed significantly higher survival after transplanting and drought stress than cuttings rooted in basal medium and treated in the same way.

  19. Adventitious Reinforcement of Maladaptive Stimulus Control Interferes with Learning.

    PubMed

    Saunders, Kathryn J; Hine, Kathleen; Hayashi, Yusuke; Williams, Dean C

    2016-09-01

    Persistent error patterns sometimes develop when teaching new discriminations. These patterns can be adventitiously reinforced, especially during long periods of chance-level responding (including baseline). Such behaviors can interfere with learning a new discrimination. They can also disrupt already learned discriminations, if they re-emerge during teaching procedures that generate errors. We present an example of this process. Our goal was to teach a boy with intellectual disabilities to touch one of two shapes on a computer screen (in technical terms, a simple simultaneous discrimination). We used a size-fading procedure. The correct stimulus was at full size, and the incorrect-stimulus size increased in increments of 10 %. Performance was nearly error free up to and including 60 % of full size. In a probe session with the incorrect stimulus at full size, however, accuracy plummeted. Also, a pattern of switching between choices, which apparently had been established in classroom instruction, re-emerged. The switching pattern interfered with already-learned discriminations. Despite having previously mastered a fading step with the incorrect stimulus up to 60 %, we were unable to maintain consistently high accuracy beyond 20 % of full size. We refined the teaching program such that fading was done in smaller steps (5 %), and decisions to "step back" to a smaller incorrect stimulus were made after every 5-instead of 20-trials. Errors were rare, switching behavior stopped, and he mastered the discrimination. This is a practical example of the importance of designing instruction that prevents adventitious reinforcement of maladaptive discriminated response patterns by reducing errors during acquisition. PMID:27622128

  20. In vitro regeneration of Salix nigra from adventitious shoots.

    PubMed

    Lyyra, Satu; Lima, Amparo; Merkle, Scott A

    2006-07-01

    Black willow (Salix nigra Marsh.) is the largest and only commercially important willow species in North America. It is a candidate for phytoremediation of polluted soils because it is fast-growing and thrives on floodplains throughout eastern USA. Our objective was to develop a protocol for the in vitro regeneration of black willow plants that could serve as target material for gene transformation. Unexpanded inflorescence explants were excised from dormant buds collected from three source trees and cultured on woody plant medium (WPM) supplemented with one of: (1) 0.1 mg l(-1) thidiazuron (TDZ); (2) 0.5 mg l(-1) 6-benzoaminopurine (BAP); or (3) 1 mg l(-1) BAP. All plant growth regulator (PGR) treatments induced direct adventitious bud formation from the genotypes. The percentage of explants producing buds ranged from 20 to 92%, depending on genotype and treatment. Although most of the TDZ-treated inflorescences produced buds, these buds failed to elongate into shoots. Buds on explants treated with BAP elongated into shoots that were easily rooted in vitro and further established in potting mix in high humidity. The PGR treatments significantly affected shoot regeneration frequency (P < 0.01). The highest shoot regeneration frequency (36%) was achieved with Genotype 3 cultured on 0.5 mg l(-1) BAP. Mean number of shoots per explant varied from one to five. The ability of black willow inflorescences to produce adventitious shoots makes them potential targets for Agrobacterium-mediated transformation with heavy-metal-resistant genes for phytoremediation. PMID:16585042

  1. Diethylnitrosamine-induced expression of germline-specific genes and pluripotency factors, including vasa and oct4, in medaka somatic cells.

    PubMed

    Shen, Jialing; Yokota, Shinpei; Yokoi, Hayato; Suzuki, Tohru

    2016-09-16

    Various methods have been developed to reprogram mammalian somatic cells into pluripotent cells as well as to directly reprogram somatic cells into other cell lineages. We are interested in applying these methods to fish, and here, we examined whether mRNA expression of germline-specific genes (vasa, nanos2, -3) and pluripotency factors (oct4, sox2, c-myc, nanog) is inducible in somatic cells of Japanese medaka (Oryzias latipes). We found that the expression of vasa is induced in the gut and regenerating fin by exposure to a carcinogen, diethylnitrosamine (DEN). Induction of vasa in the gut started on the 5th day of treatment with >50 ppm DEN. In addition, nanos2, -3, oct4, sox2, klf4, c-myc, and nanog were also expressed simultaneously in some vasa-positive gut and regenerating fin samples. Vasa-positive cells were detected by immunohistochemistry (IHC) in the muscle surrounding the gut and in the wound epidermis, blastema, and fibroblast-like cells in regenerating fin. In vasa:GFP transgenic medaka, green fluorescent protein (GFP) fluorescence appeared in the wound epidermis and fibroblast-like cells in the regenerating fin following DEN exposure, in agreement with the IHC data. Our data show that mRNA expression of genes relevant to germ cell specification and pluripotency can be induced in fish somatic cells by exposure to DEN, suggesting the possibility of efficient and rapid cell reprogramming of fish somatic cells. PMID:27514449

  2. Adventitial fibroblasts are activated in the early stages of atherosclerosis in the apolipoprotein E knockout mouse

    SciTech Connect

    Xu Fang; Ji Jian; Li Li; Chen Rong; Hu Weicheng . E-mail: huweicheng@sdu.edu.cn

    2007-01-19

    The role of the adventitia in vascular function and vascular lesion formation has been largely ignored. This study observed the activation of the adventitia and specifically the fibroblasts in the development of atherosclerosis in the apoE(-/-) mouse. The results showed a gradual increase in expression of collagen types I and III after 2, 4, and 8 weeks of hyperlipidic diet. The earliest expression of monocyte chemoattractant protein-1 (MCP-1) protein and mRNA was detected in the adventitial fibroblast before the formation of intimal lesions. Proliferation, too, was first found in the adventitial fibroblasts. We hypothesize that the adventitial fibroblast is activated in the early stage of atherosclerosis. Adventitial inflammation may be an early event in the development of atherosclerotic lesions.

  3. Adventitial gene transfer of catalase attenuates angiotensin II-induced vascular remodeling.

    PubMed

    Liu, Cun-Fei; Zhang, Jia; Shen, Kai; Gao, Ping-Jin; Wang, Hai-Ya; Jin, Xin; Meng, Chao; Fang, Ning-Yuan

    2015-04-01

    Vascular adventitia and adventitia‑derived reactive oxygen species (ROS) contribute to vascular remodeling following vascular injury. A previous ex vivo study in adventitial fibroblasts showed that catalase, one of most important anti‑oxide enzymes, was downregulated by angiotensin II (AngII). The aim of the present study was to investigate whether adventitial gene transfer of catalase affects AngII‑induced vascular remodeling in vivo. Adenoviruses co‑expressing catalase and enhanced green fluorescent protein (eGFP) or expressing eGFP only were applied to the adventitial surface of common carotid arteries of Sprague‑Dawley rats. Alzet minipumps administering AngII (0.75 mg/kg/day) were then implanted subcutaneously for 14 days. Systolic blood pressure and biological parameters of vascular remodeling were measured in each group. Adventitial fibroblasts were cultured and p38 mitogen‑activated protein kinase (MAPK) phosphorylation was measured using western blot analysis. The results showed that adventitial gene transfer of catalase had no effect on AngII‑induced systolic blood pressure elevation. However, catalase adenovirus transfection significantly inhibited AngII‑induced media hypertrophy compared with that of the control virus (P<0.05). In addition, catalase transfection significantly attenuated AngII‑induced ROS generation, macrophage infiltration, collagen deposition and adventitial α‑smooth muscle actin expression. Furthermore, catalase transfection significantly inhibited the AngII‑induced increase in p38MAPK phosphorylation. In conclusion, the results of the present study demonstrated that adventitial gene transfer of catalase significantly attenuated AngII‑induced vascular remodeling in rats via inhibition of adventitial p38MAPK phosphorylation.

  4. Somatic stem cells express Piwi and Vasa genes in an adult ctenophore: ancient association of "germline genes" with stemness.

    PubMed

    Alié, Alexandre; Leclère, Lucas; Jager, Muriel; Dayraud, Cyrielle; Chang, Patrick; Le Guyader, Hervé; Quéinnec, Eric; Manuel, Michaël

    2011-02-01

    Stem cells are essential for animal development and adult tissue homeostasis, and the quest for an ancestral gene fingerprint of stemness is a major challenge for evolutionary developmental biology. Recent studies have indicated that a series of genes, including the transposon silencer Piwi and the translational activator Vasa, specifically involved in germline determination and maintenance in classical bilaterian models (e.g., vertebrates, fly, nematode), are more generally expressed in adult multipotent stem cells in other animals like flatworms and hydras. Since the progeny of these multipotent stem cells includes both somatic and germinal derivatives, it remains unclear whether Vasa, Piwi, and associated genes like Bruno and PL10 were ancestrally linked to stemness, or to germinal potential. We have investigated the expression of Vasa, two Piwi paralogues, Bruno and PL10 in Pleurobrachia pileus, a member of the early-diverging phylum Ctenophora, the probable sister group of cnidarians. These genes were all expressed in the male and female germlines, and with the exception of one of the Piwi paralogues, they showed similar expression patterns within somatic territories (tentacle root, comb rows, aboral sensory complex). Cytological observations and EdU DNA-labelling and long-term retention experiments revealed concentrations of stem cells closely matching these gene expression areas. These stem cell pools are spatially restricted, and each specialised in the production of particular types of somatic cells. These data unveil important aspects of cell renewal within the ctenophore body and suggest that Piwi, Vasa, Bruno, and PL10 belong to a gene network ancestrally acting in two distinct contexts: (i) the germline and (ii) stem cells, whatever the nature of their progeny.

  5. Aquaporin-1 water channels in short and long loop descending thin limbs and in descending vasa recta in rat kidney.

    PubMed

    Nielsen, S; Pallone, T; Smith, B L; Christensen, E I; Agre, P; Maunsbach, A B

    1995-06-01

    The localization of aquaporin-1 water channels (AQP-1) in nephron and vascular structures in rat kidney were characterized, because vascular bundles are known to play a key role in urinary concentration. Immunohistochemistry and immunoelectron microscopy were applied on thin cryosections or ultrathin Lowicryl sections, using an optimized freeze-substitution method. Within the vascular bundles, AQP-1 is localized in descending thin limbs (DTL) of short nephrons in apical and basolateral membranes. The expression in DTL of short nephrons is considerably lower compared with the expression in long nephrons, consistent with the known lower osmotic water permeability of this segment. Furthermore, DTL of short nephrons expressing AQP-1 continue abruptly into a thin limb segment without AQP-1. This suggests the existence of a novel thin limb epithelium in the outer medulla. Extensive expression of AQP-1 is observed in apical and basolateral membranes of DTL of long nephrons, which are localized in the periphery of the vascular bundles. The expression decreases along the axis of long nephron DTLs in correlation with the known water permeability characteristics of thin limb segments. DTLs of both short and long nephrons continue abruptly into thin limb segments without AQP-1 expression, revealing an abrupt cell-to-cell transition. In vasa recta, AQP-1 is selectively localized in the nonfenestrated endothelium of descending vasa recta, whereas the fenestrated endothelium of ascending vesa recta and peritubular capillaries do not express AQP-1. AQP-1 is localized in both apical and basolateral plasma membranes, which is logical for transendothelial water transport. Isolated perfused descending vasa recta display high water permeability, and, unlike sodium permeability, diffusional water permeability is partly inhibited by mercurials, thus substantiating the presence of mercurial-sensitive water channels in descending vasa recta. Thus AQP-1 is localized in DTL and descending

  6. Adventitious root induction in Arabidopsis thaliana as a model for in vitro root organogenesis.

    PubMed

    Verstraeten, Inge; Beeckman, Tom; Geelen, Danny

    2013-01-01

    Adventitious root formation, the development of roots on non-root tissue (e.g. leaves, hypocotyls and stems) is a critical step during micropropagation. Although root induction treatments are routinely used for a large number of species micropropagated in vitro as well as for in vivo cuttings, the mechanisms controlling adventitious rooting are still poorly understood. Researchers attempt to gain better insight into the molecular aspects by studying adventitious rooting in Arabidopsis thaliana. The existing assay involves etiolation of seedlings and measurements of de novo formed roots on the elongated hypocotyl. The etiolated hypocotyls express a novel auxin-controlled signal transduction pathway in which auxin response factors (ARFs), microRNAs and environmental conditions that drive adventitious rooting are integrated. An alternative assay makes use of so-called thin cell layers (TCL), excised strips of cells from the inflorescence stem of Arabidopsis thaliana. However, both the etiolated seedling system and the TCL assay are only distantly related to industrial rooting processes in which roots are induced on adult stem tissue. Here, we describe an adventitious root induction system that uses segments of the inflorescence stems of Arabidopsis thaliana, which have a histological structure similar to cuttings or in vitro micropropagated shoots. The system allows multiple treatments with chemicals as well as the evaluation of different environmental conditions on a large number of explants. It is therefore suitable for high throughput chemical screenings and experiments that require numerous data points for statistical analysis. Using this assay, the adventitious root induction capacity of classical auxins was evaluated and a differential response to the different auxins could be demonstrated. NAA, IBA and IAA stimulated adventitious rooting on the stem segment, whereas 2,4-D and picloram did not. Light conditions profoundly influenced the root induction capacity

  7. DDX4 (VASA) is conserved in germ cell development in marsupials and monotremes.

    PubMed

    Hickford, Danielle E; Frankenberg, Stephen; Pask, Andrew J; Shaw, Geoff; Renfree, Marilyn B

    2011-10-01

    DDX4 (VASA) is an RNA helicase expressed in the germ cells of all animals. To gain greater insight into the role of this gene in mammalian germ cell development, we characterized DDX4 in both a marsupial (the tammar wallaby) and a monotreme (the platypus). DDX4 is highly conserved between eutherian, marsupial, and monotreme mammals. DDX4 protein is absent from tammar fetal germ cells but is present from Day 1 postpartum in both sexes. The distribution of DDX4 protein during oogenesis and spermatogenesis in the tammar is similar to eutherians. Female tammar germ cells contain DDX4 protein throughout all stages of postnatal oogenesis. In males, DDX4 is in gonocytes, and during spermatogenesis it is present in spermatocytes and round spermatids. A similar distribution of DDX4 occurs in the platypus during spermatogenesis. There are several DDX4 isoforms in the tammar, resulting from both pre- and posttranslational modifications. DDX4 in marsupials and monotremes has multiple splice variants and polyadenylation motifs. Using in silico analyses of genomic databases, we found that these previously unreported splice variants also occur in eutherians. In addition, several elements implicated in the control of Ddx4 expression in the mouse, including RGG (arginine-glycine-glycine) and dimethylation of arginine motifs and CpG islands within the Ddx4 promoter, are also highly conserved. Collectively these data suggest that DDX4 is essential for the regulation of germ cell proliferation and differentiation across all three extant mammalian groups-eutherians, marsupials, and monotremes.

  8. Dissecting the contribution of microtubule behaviour in adventitious root induction

    PubMed Central

    Abu-Abied, Mohamad; Rogovoy (Stelmakh), Oksana; Mordehaev, Inna; Grumberg, Marina; Elbaum, Rivka; Wasteneys, Geoffrey O.; Sadot, Einat

    2015-01-01

    Induction of adventitious roots (ARs) in recalcitrant plants often culminates in cell division and callus formation rather than root differentiation. Evidence is provided here to suggest that microtubules (MTs) play a role in the shift from cell division to cell differentiation during AR induction. First, it was found that fewer ARs form in the temperature-sensitive mutant mor1-1, in which the MT-associated protein MOR1 is mutated, and in bot1-1, in which the MT-severing protein katanin is mutated. In the two latter mutants, MT dynamics and form are perturbed. By contrast, the number of ARs increased in RIC1-OX3 plants, in which MT bundling is enhanced and katanin is activated. In addition, any1 plants in which cell walls are perturbed made more ARs than wild-type plants. MT perturbations during AR induction in mor1-1 or in wild-type hypocotyls treated with oryzalin led to the formation of amorphous clusters of cells reminiscent of callus. In these cells a specific pattern of polarized light retardation by the cell walls was lost. PIN1 polarization and auxin maxima were hampered and differentiation of the epidermis was inhibited. It is concluded that a fine-tuned crosstalk between MTs, cell walls, and auxin transport is required for proper AR induction. PMID:25788735

  9. Adventitious rooting declines with the vegetative to reproductive switch and involves a changed auxin homeostasis

    PubMed Central

    Rasmussen, Amanda; Hosseini, Seyed Abdollah; Hajirezaei, Mohammed-Reza; Druege, Uwe; Geelen, Danny

    2015-01-01

    Adventitious rooting, whereby roots form from non-root tissues, is critical to the forestry and horticultural industries that depend on propagating plants from cuttings. A major problem is that age of the tissue affects the ability of the cutting to form adventitious roots. Here, a model system has been developed using Pisum sativum to differentiate between different interpretations of ageing. It is shown that the decline in adventitious rooting is linked to the ontogenetic switch from vegetative to floral and is mainly attributed to the cutting base. Using rms mutants it is demonstrated that the decline is not a result of increased strigolactones inhibiting adventitious root formation. Monitoring endogenous levels of a range of other hormones including a range of cytokinins in the rooting zone revealed that a peak in jasmonic acid is delayed in cuttings from floral plants. Additionally, there is an early peak in indole-3-acetic acid levels 6h post excision in cuttings from vegetative plants, which is absent in cuttings from floral plants. These results were confirmed using DR5:GUS expression. Exogenous supplementation of young cuttings with either jasmonic acid or indole-3-acetic acid promoted adventitious rooting, but neither of these hormones was able to promote adventitious rooting in mature cuttings. DR5:GUS expression was observed to increase in juvenile cuttings with increasing auxin treatment but not in the mature cuttings. Therefore, it seems the vegetative to floral ontogenetic switch involves an alteration in the tissue’s auxin homeostasis that significantly reduces the indole-3-acetic acid pool and ultimately results in a decline in adventitious root formation. PMID:25540438

  10. Adventitious rooting declines with the vegetative to reproductive switch and involves a changed auxin homeostasis.

    PubMed

    Rasmussen, Amanda; Hosseini, Seyed Abdollah; Hajirezaei, Mohammed-Reza; Druege, Uwe; Geelen, Danny

    2015-03-01

    Adventitious rooting, whereby roots form from non-root tissues, is critical to the forestry and horticultural industries that depend on propagating plants from cuttings. A major problem is that age of the tissue affects the ability of the cutting to form adventitious roots. Here, a model system has been developed using Pisum sativum to differentiate between different interpretations of ageing. It is shown that the decline in adventitious rooting is linked to the ontogenetic switch from vegetative to floral and is mainly attributed to the cutting base. Using rms mutants it is demonstrated that the decline is not a result of increased strigolactones inhibiting adventitious root formation. Monitoring endogenous levels of a range of other hormones including a range of cytokinins in the rooting zone revealed that a peak in jasmonic acid is delayed in cuttings from floral plants. Additionally, there is an early peak in indole-3-acetic acid levels 6h post excision in cuttings from vegetative plants, which is absent in cuttings from floral plants. These results were confirmed using DR5:GUS expression. Exogenous supplementation of young cuttings with either jasmonic acid or indole-3-acetic acid promoted adventitious rooting, but neither of these hormones was able to promote adventitious rooting in mature cuttings. DR5:GUS expression was observed to increase in juvenile cuttings with increasing auxin treatment but not in the mature cuttings. Therefore, it seems the vegetative to floral ontogenetic switch involves an alteration in the tissue's auxin homeostasis that significantly reduces the indole-3-acetic acid pool and ultimately results in a decline in adventitious root formation. PMID:25540438

  11. Lipopolysaccharide promotes lipid accumulation in human adventitial fibroblasts via TLR4-NF-κB pathway

    PubMed Central

    2012-01-01

    Background Atherosclerosis is a chronic degenerative disease of the arteries and is thought to be one of the most common causes of death globally. In recent years, the functions of adventitial fibroblasts in the development of atherosclerosis and tissue repair have gained increased interests. LPS can increase the morbidity and mortality of atherosclerosis-associated cardiovascular disease. Although LPS increases neointimal via TLR4 activation has been reported, how LPS augments atherogenesis through acting on adventitial fibroblasts is still unknown. Here we explored lipid deposition within adventitial fibroblasts mediated by lipopolysaccharide (LPS) to imitate inflammatory conditions. Results In our study, LPS enhanced lipid deposition by the up-regulated expression of adipose differentiation-related protein (ADRP) as the silencing of ADRP abrogated lipid deposition in LPS-activated adventitial fibroblasts. In addition, pre-treatment with anti-Toll-like receptor 4 (TLR4) antibody diminished the LPS-induced lipid deposition and ADRP expression. Moreover, LPS induced translocation of nuclear factor-κB (NF-κB), which could markedly up-regulate lipid deposition as pre-treatment with the NF-κB inhibitor, PDTC, significantly reduced lipid droplets. In addition, the lowering lipid accumulation was accompanied with the decreased ADRP expression. Furthermore, LPS-induced adventitial fibroblasts secreted more monocyte chemoattractant protein (MCP-1), compared with transforming growth factor-β1 (TGF-β1). Conclusions Taken together, these results suggest that LPS promotes lipid accumulation via the up-regulation of ADRP expression through TLR4 activated downstream of NF-κB in adventitial fibroblasts. Increased levels of MCP-1 released from LPS-activated adventitial fibroblasts and lipid accumulation may accelerate monocytes recruitment and lipid-laden macrophage foam cells formation. Here, our study provides a new explanation as to how bacterial infection contributes to

  12. Identification of new adventitious rooting mutants amongst suppressors of the Arabidopsis thaliana superroot2 mutation

    PubMed Central

    Pacurar, Daniel Ioan; Pacurar, Monica Lacramioara; Schwambach, Joseli; Bellini, Catherine

    2014-01-01

    The plant hormone auxin plays a central role in adventitious rooting and is routinely used with many economically important, vegetatively propagated plant species to promote adventitious root initiation and development on cuttings. Nevertheless the molecular mechanisms through which it acts are only starting to emerge. The Arabidopsis superroot2-1 (sur2-1) mutant overproduces auxin and, as a consequence, develops excessive adventitious roots in the hypocotyl. In order to increase the knowledge of adventitious rooting and of auxin signalling pathways and crosstalk, this study performed a screen for suppressors of superroot2-1 phenotype. These suppressors provide a new resource for discovery of genetic players involved in auxin signalling pathways or at the crosstalk of auxin and other hormones or environmental signals. This study reports the identification and characterization of 26 sur2-1 suppressor mutants, several of which were identified as mutations in candidate genes involved in either auxin biosynthesis or signalling. In addition to confirming the role of auxin as a central regulator of adventitious rooting, superroot2 suppressors indicated possible crosstalk with ethylene signalling in this process. PMID:24596172

  13. DDX4 (VASA) is conserved in germ cell development in marsupials and monotremes.

    PubMed

    Hickford, Danielle E; Frankenberg, Stephen; Pask, Andrew J; Shaw, Geoff; Renfree, Marilyn B

    2011-10-01

    DDX4 (VASA) is an RNA helicase expressed in the germ cells of all animals. To gain greater insight into the role of this gene in mammalian germ cell development, we characterized DDX4 in both a marsupial (the tammar wallaby) and a monotreme (the platypus). DDX4 is highly conserved between eutherian, marsupial, and monotreme mammals. DDX4 protein is absent from tammar fetal germ cells but is present from Day 1 postpartum in both sexes. The distribution of DDX4 protein during oogenesis and spermatogenesis in the tammar is similar to eutherians. Female tammar germ cells contain DDX4 protein throughout all stages of postnatal oogenesis. In males, DDX4 is in gonocytes, and during spermatogenesis it is present in spermatocytes and round spermatids. A similar distribution of DDX4 occurs in the platypus during spermatogenesis. There are several DDX4 isoforms in the tammar, resulting from both pre- and posttranslational modifications. DDX4 in marsupials and monotremes has multiple splice variants and polyadenylation motifs. Using in silico analyses of genomic databases, we found that these previously unreported splice variants also occur in eutherians. In addition, several elements implicated in the control of Ddx4 expression in the mouse, including RGG (arginine-glycine-glycine) and dimethylation of arginine motifs and CpG islands within the Ddx4 promoter, are also highly conserved. Collectively these data suggest that DDX4 is essential for the regulation of germ cell proliferation and differentiation across all three extant mammalian groups-eutherians, marsupials, and monotremes. PMID:21653890

  14. Effect of sodium chloride gradients on water flux in rat descending vasa recta.

    PubMed Central

    Pallone, T L

    1991-01-01

    In the hydropenic kidney, volume efflux from descending vasa recta (DVR) occurs despite an intracapillary oncotic pressure that exceeds hydraulic pressure. That finding has been attributed to small solute gradients which may provide an additional osmotic driving force favoring water transport from DVR plasma to the papillary interstitium. To test this hypothesis, axial gradients of NaCl and urea in the papilla were eliminated by administration of furosemide and saline. DVR were then blocked with paraffin and microperfused at 10 nl/min with a buffer containing albumin, fluorescein isothiocyanate labeled dextran (FITC-Dx), 22Na, and NaCl in a concentration of 0 (hypotonic to the interstitium), 161 (isotonic) or 322 mM (hypertonic). Collectate was obtained from the perfused DVR by micropuncture and the collectate-to-perfusate ratios of FITC-Dx and 22Na were measured. A mathematical model was employed to determine DVR permeability (Ps) and reflection coefficient to NaCl (sigma NaCl). The rate of transport of water from the DVR lumen to the papillary interstitium was 2.8 +/- 0.3 (Nv = 22), -0.19 +/- 0.4 (Nv = 15), and -2.3 +/- 0.3 nl/min (Nv = 21) (mean +/- SE) when perfusate NaCl was 0, 161, or 322 mM, respectively (Nv = number of DVR perfused). The collectate-to-perfusate 22Na concentration ratios were 0.34 +/- 0.04, 0.36 +/- 0.04 and 0.37 +/- 0.03 for those groups, respectively. Based on these data, Ps is calculated to be 60.4 x 10(-5) +/- 4.0 x 10(-5) cm/s and sigma NaCl less than 0.05. The results of this study confirm that transcapillary NaCl concentrations gradients induce water movement across the wall of the DVR. PMID:1985089

  15. Adventitious agents in viral vaccines: lessons learned from 4 case studies.

    PubMed

    Petricciani, John; Sheets, Rebecca; Griffiths, Elwyn; Knezevic, Ivana

    2014-09-01

    Since the earliest days of biological product manufacture, there have been a number of instances where laboratory studies provided evidence for the presence of adventitious agents in a marketed product. Lessons learned from such events can be used to strengthen regulatory preparedness for the future. We have therefore selected four instances where an adventitious agent, or a signal suggesting the presence of an agent, was found in a viral vaccine, and have developed a case study for each. The four cases are: a) SV40 in polio vaccines; b) bacteriophage in measles and polio vaccines; c) reverse transcriptase in measles and mumps vaccines; and d) porcine circovirus and porcine circovirus DNA sequences in rotavirus vaccines. The lessons learned from each event are discussed. Based in part on those experiences, certain scientific principles have been identified by WHO that should be considered in regulatory risk evaluation if an adventitious agent is found in a marketed vaccine in the future.

  16. Ectopic expression of Cvh (Chicken Vasa homologue) mediates the reprogramming of chicken embryonic stem cells to a germ cell fate.

    PubMed

    Lavial, Fabrice; Acloque, Hervé; Bachelard, Elodie; Nieto, M Angela; Samarut, Jacques; Pain, Bertrand

    2009-06-01

    When they are derived from blastodermal cells of the pre-primitive streak in vitro, the pluripotency of Chicken Embryonic Stem Cells (cESC) can be controlled by the cPouV and Nanog genes. These cESC can differentiate into derivatives of the three germ layers both in vitro and in vivo, but they only weakly colonize the gonads of host embryos. By contrast, non-cultured blastodermal cells and long-term cultured chicken primordial germ cells maintain full germline competence. This restriction in the germline potential of the cESC may result from either early germline determination in the donor embryos or it may occur as a result of in vitro culture. We are interested in understanding the genetic determinants of germline programming. The RNA binding protein Cvh (Chicken Vasa Homologue) is considered as one such determinant, although its role in germ cell physiology is still unclear. Here we show that the exogenous expression of Cvh, combined with appropriate culture conditions, induces cESC reprogramming towards a germ cell fate. Indeed, these cells express the Dazl, Tudor and Sycp3 germline markers, and they display improved germline colonization and adopt a germ cell fate when injected into recipient embryos. Thus, our results demonstrate that Vasa can drive ES cell differentiation towards the germ cell lineage, both in vitro and in vivo.

  17. Distinct effects of auxin and light on adventitious root development in Eucalyptus saligna and Eucalyptus globulus.

    PubMed

    Fett-Neto, A G; Fett, J P; Veira Goulart, L W; Pasquali, G; Termignoni, R R; Ferreira, A G

    2001-05-01

    Adventitious rooting is essential for vegetative propagation of woody species. We studied the effects of auxin and light on the development of adventitious roots in cuttings obtained from seedlings of Eucalyptus saligna Smith and E. globulus Labill in an attempt to characterize the adventitious rooting process and identify factors controlling rhizogenesis. Root development was scored as rooting percentage, root density (roots per rooted cutting), mean rooting time and root length. In both species, rooting time was reduced in the presence of auxin. Cuttings from 2-month-old E. saligna seedlings were responsive to lower auxin concentrations than comparable cuttings from E. globulus seedlings. Cuttings from 3-month-old E. saligna seedlings rooted promptly and rooting was not significantly affected by light conditions. In contrast, rooting of cuttings from 3-month-old E. globulus seedlings exhibited recalcitrant behavior and no roots were formed if illuminated during the root formation phase. Effective root regeneration of E. globulus cuttings was obtained by a 4-day exposure to 10 mg l(-1) IBA and culture in darkness during the root formation step. Loss of rooting capacity with seedling age was more pronounced in E. globulus than in E. saligna. The possibility of switching adventitious rooting off and on by manipulating light regime and exogenous auxin supply in E. globulus, and the constitutive nature of rooting in E. saligna may provide useful models for examining the rooting process at the biochemical and molecular levels in Eucalyptus. PMID:11340046

  18. Tissue sealing device associated thermal spread: a comparison of histologic methods for detecting adventitial collagen denaturation

    NASA Astrophysics Data System (ADS)

    Jones, Ryan M.; Grisez, Brian T.; Thomas, Aaron C.; Livengood, Ryan H.; Coad, James E.

    2013-02-01

    Thermal spread (thermal tissue damage) results from heat conduction through the tissues immediately adjacent to a hyperthermic tissue sealing device. The extent of such heat conduction can be assessed by the detection of adventitial collagen denaturation. Several histologic methods have been reported to measure adventitial collagen denaturation as a marker of thermal spread. This study compared hematoxylin and eosin staining, Gomori trichrome staining and loss of collagen birefringence for the detection of collagen denaturation. Twenty-eight ex vivo porcine carotid arteries were sealed with a commercially available, FDA-approved tissue sealing device. Following formalin fixation and paraffin embedding, two 5-micron tissue sections were hematoxylin and eosin and Gomori trichrome stained. The hematoxylin and eosin-stained section was evaluated by routine bright field microscopy and under polarized light. The trichromestained section was evaluated by routine bright field microscopy. Radial and midline adventitial collagen denaturation measurements were made for both the top and bottom jaw sides of each seal. The adventitial collagen denaturation lengths were determined using these three methods and statistically compared. The results showed that thermal spread, as represented by histologically detected collagen denaturation, is technique dependent. In this study, the trichrome staining method detected significantly less thermal spread than the hematoxylin and eosin staining and birefringence methods. Of the three methods, hematoxylin and eosin staining provided the most representative results for true thermal spread along the adjacent artery.

  19. Early steps of adventitious rooting: morphology, hormonal profiling and carbohydrate turnover in carnation stem cuttings.

    PubMed

    Agulló-Antón, María Ángeles; Ferrández-Ayela, Almudena; Fernández-García, Nieves; Nicolás, Carlos; Albacete, Alfonso; Pérez-Alfocea, Francisco; Sánchez-Bravo, José; Pérez-Pérez, José Manuel; Acosta, Manuel

    2014-03-01

    The rooting of stem cuttings is a common vegetative propagation practice in many ornamental species. A detailed analysis of the morphological changes occurring in the basal region of cultivated carnation cuttings during the early stages of adventitious rooting was carried out and the physiological modifications induced by exogenous auxin application were studied. To this end, the endogenous concentrations of five major classes of plant hormones [auxin, cytokinin (CK), abscisic acid, salicylic acid (SA) and jasmonic acid] and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid were analyzed at the base of stem cuttings and at different stages of adventitious root formation. We found that the stimulus triggering the initiation of adventitious root formation occurred during the first hours after their excision from the donor plant, due to the breakdown of the vascular continuum that induces auxin accumulation near the wounding. Although this stimulus was independent of exogenously applied auxin, it was observed that the auxin treatment accelerated cell division in the cambium and increased the sucrolytic activities at the base of the stem, both of which contributed to the establishment of the new root primordia at the stem base. Further, several genes involved in auxin transport were upregulated in the stem base either with or without auxin application, while endogenous CK and SA concentrations were specially affected by exogenous auxin application. Taken together our results indicate significant crosstalk between auxin levels, stress hormone homeostasis and sugar availability in the base of the stem cuttings in carnation during the initial steps of adventitious rooting. PMID:24117983

  20. Early steps of adventitious rooting: morphology, hormonal profiling and carbohydrate turnover in carnation stem cuttings.

    PubMed

    Agulló-Antón, María Ángeles; Ferrández-Ayela, Almudena; Fernández-García, Nieves; Nicolás, Carlos; Albacete, Alfonso; Pérez-Alfocea, Francisco; Sánchez-Bravo, José; Pérez-Pérez, José Manuel; Acosta, Manuel

    2014-03-01

    The rooting of stem cuttings is a common vegetative propagation practice in many ornamental species. A detailed analysis of the morphological changes occurring in the basal region of cultivated carnation cuttings during the early stages of adventitious rooting was carried out and the physiological modifications induced by exogenous auxin application were studied. To this end, the endogenous concentrations of five major classes of plant hormones [auxin, cytokinin (CK), abscisic acid, salicylic acid (SA) and jasmonic acid] and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid were analyzed at the base of stem cuttings and at different stages of adventitious root formation. We found that the stimulus triggering the initiation of adventitious root formation occurred during the first hours after their excision from the donor plant, due to the breakdown of the vascular continuum that induces auxin accumulation near the wounding. Although this stimulus was independent of exogenously applied auxin, it was observed that the auxin treatment accelerated cell division in the cambium and increased the sucrolytic activities at the base of the stem, both of which contributed to the establishment of the new root primordia at the stem base. Further, several genes involved in auxin transport were upregulated in the stem base either with or without auxin application, while endogenous CK and SA concentrations were specially affected by exogenous auxin application. Taken together our results indicate significant crosstalk between auxin levels, stress hormone homeostasis and sugar availability in the base of the stem cuttings in carnation during the initial steps of adventitious rooting.

  1. Nitric oxide is required for hydrogen gas-induced adventitious root formation in cucumber.

    PubMed

    Zhu, Yongchao; Liao, Weibiao; Wang, Meng; Niu, Lijuan; Xu, Qingqing; Jin, Xin

    2016-05-20

    Hydrogen gas (H2) is involved in plant development and stress responses. Cucumber explants were used to study whether nitric oxide (NO) is involved in H2-induced adventitious root development. The results revealed that 50% and 100% hydrogen-rich water (HRW) apparently promoted the development of adventitious root in cucumber. While, the responses of HRW-induced adventitious rooting were blocked by a specific NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO), NO synthase (NOS) enzyme inhibitor N(G)-nitro-l-arginine methylester hydrochloride (l-NAME) and nitrate reductase (NR) inhibitor NaN3. HRW also increased NO content and NOS and NR activity both in a dose- and time-dependent fashion. Moreover, molecular evidence showed that HRW up-regulated NR genes expression in explants. The results indicate the importance of NOS and NR enzymes, which might be responsible for NO production in explants during H2-induced root organogenesis. Additionally, peroxidase (POD) and indoleacetic acid oxidase (IAAO) activity was significantly decreased in the explants treated with HRW, while HRW treatment significantly increased polyphenol oxidase (PPO) activity. In addition, cPTIO, l-NAME and NaN3 inhibited the actions of HRW on the activity of these enzymes. Together, NO may be involved in H2-induced adventitious rooting, and NO may be acting downstream in plant H2 signaling cascade. PMID:27010347

  2. Adventitial cystic disease of the common femoral vein presenting as deep vein thrombosis.

    PubMed

    Kim, Young-Kyun; Chun, Ho Jong; Hwang, Jeong Kye; Kim, Ji Il; Kim, Sang Dong; Park, Sun-Cheol; Moon, In Sung

    2016-07-01

    Adventitial cystic disease of the common femoral vein is a rare condition. We herein report the case of a 50-year-old woman who presented with painless swelling in her left lower leg that resembled deep vein thrombosis. She underwent femoral exploration and excision of the cystic wall. The presentation, investigation, treatment, and pathology of this condition are discussed with a literature review.

  3. Sympathetic nerve-derived ATP regulates renal medullary vasa recta diameter via pericyte cells: a role for regulating medullary blood flow?

    PubMed

    Crawford, C; Wildman, S S P; Kelly, M C; Kennedy-Lydon, T M; Peppiatt-Wildman, C M

    2013-01-01

    Pericyte cells are now known to be a novel locus of blood flow control, being able to regulate capillary diameter via their unique morphology and expression of contractile proteins. We have previously shown that exogenous ATP causes constriction of vasa recta via renal pericytes, acting at a variety of membrane bound P2 receptors on descending vasa recta (DVR), and therefore may be able to regulate medullary blood flow (MBF). Regulation of MBF is essential for appropriate urine concentration and providing essential oxygen and nutrients to this region of high, and variable, metabolic demand. Various sources of endogenous ATP have been proposed, including from epithelial, endothelial, and red blood cells in response to stimuli such as mechanical stimulation, local acidosis, hypoxia, and exposure to various hormones. Extensive sympathetic innervation of the nephron has previously been shown, however the innervation reported has focused around the proximal and distal tubules, and ascending loop of Henle. We hypothesize that sympathetic nerves are an additional source of ATP acting at renal pericytes and therefore regulate MBF. Using a rat live kidney slice model in combination with video imaging and confocal microscopy techniques we firstly show sympathetic nerves in close proximity to vasa recta pericytes in both the outer and inner medulla. Secondly, we demonstrate pharmacological stimulation of sympathetic nerves in situ (by tyramine) evokes pericyte-mediated vasoconstriction of vasa recta capillaries; inhibited by the application of the P2 receptor antagonist suramin. Lastly, tyramine-evoked vasoconstriction of vasa recta by pericytes is significantly less than ATP-evoked vasoconstriction. Sympathetic innervation may provide an additional level of functional regulation in the renal medulla that is highly localized. It now needs to be determined under which physiological/pathophysiological circumstances that sympathetic innervation of renal pericytes is important

  4. Cloning, characterization, and expression analysis of the DEAD-box family genes, Fc-vasa and Fc-PL10a, in Chinese shrimp ( Fenneropenaeus chinensis)

    NASA Astrophysics Data System (ADS)

    Zhou, Qianru; Shao, Mingyu; Qin, Zhenkui; Kyoung, Ho Kang; Zhang, Zhifeng

    2010-01-01

    RNA helicases of the DEAD-box and related families are involved in various cellular processes including DNA replication, DNA repair, and RNA processing. However, the function of DEAD-box proteins in aquaculture species is poorly understood at molecular level. We obtained the full-length cDNA sequences of two genes encoding helicase-related proteins, Fc-vasa and Fc-PL10a, from the testes of Chinese shrimp, Fenneropenaeus chinensis. The two predicted amino acid sequences contain all the conserved motifs characterized by the DEAD-box family and several RGG repeats in the N-terminal regions. Homology and phylogenetic analyses indicate that they belong to the vasa and PL10 subfamilies. The three-dimensional structures of the two proteins were predicted with a homology modeling approach. Both core proteins consist of two tandem RecA-like domains similar to those of the DEAD-box RNA helicase. Using reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR we found that Fc-vasa was expressed specifically in the adult gonads. Transcription decreased in the ovary but increased in the testis during gonadal development. Fc-PL10a expression was widely distributed in the tissues we examined. Using in situ hybridization, we demonstrated that the Fc-vasa transcript is localized to the cytoplasm of the spermatogonia and oocytes. Thus, our results suggest that Fc-vasa plays an important role in germ-line development, and has utility as a germ cell lineage marker which will help to generate new insight into the origin and differentiation of germ cells as well as the regulation of reproduction in F. chinensis.

  5. Expression Pattern of Mouse Vasa Homologue (MVH) in the Ovaries of C57BL/6 Female Mice

    PubMed Central

    Song, Kunkun; Ma, Wenwen; Huang, Cong; Ding, Jiahui; Cui, Dandan; Zhang, Mingmin

    2016-01-01

    Background Vasa (a DEAD-box helicase, also known as Ddx4) is an ATP-dependent RNA helicase highly conserved among all animals. Research on the presence and function of DDX4 in female mammals is limited. To gain greater insight into its distribution and role in female mice, we detected the expression of DDX4 protein in the ovaries and analyzed its expression pattern. Material/Methods MVH was detected in the cytoplasm of oocytes in all non-apoptotic follicles. Results In the present study, we found that higher expression levels of ~55–60 kDa MVH isoform in the ovaries were followed by the accumulations of preovulatory follicles. Conclusions Higher levels of MVH protein in the ovaries might prepare oocytes for the competence to resume meiosis. PMID:27460133

  6. In mouse oocytes the mitochondrion-originated germinal body-like structures accumulate mouse Vasa homologue (MVH) protein.

    PubMed

    Reunov, Arkadiy A; Reunova, Yulia A

    2015-08-01

    Mouse Vasa homologue (MVH) antibodies were applied to mouse Graafian oocytes to clarify if mitochondrion-originated germinal body-like structures, described previously by conventional electron microscopy, were associated with the germ plasm. It was found that both the mitochondrion-like structures with cristae and the germinal body-like structures that lacked any signs of cristae were labelled specifically by the anti-MVH antibody. Moreover, some granules were MVH-positive ultrastructural hybrids of the mitochondria and germinal body-like structures, the presence of which clearly supported the idea of a mitochondrial origin for the germinal body-like structures. This finding is the first evidence that mitochondrion-originated germinal body-like granules represent mouse germ plasm.

  7. Splice variants and promoter methylation status of the Bovine Vasa Homology (Bvh) gene may be involved in bull spermatogenesis

    PubMed Central

    2013-01-01

    Background Vasa is a member of the DEAD-box protein family that plays an indispensable role in mammalian spermatogenesis, particularly during meiosis. Bovine vasa homology (Bvh) of Bos taurus has been reported, however, its function in bovine testicular tissue remains obscure. This study aimed to reveal the functions of Bvh and to determine whether Bvh is a candidate gene in the regulation of spermatogenesis in bovine, and to illustrate whether its transcription is regulated by alternative splicing and DNA methylation. Results Here we report the molecular characterization, alternative splicing pattern, expression and promoter methylation status of Bvh. The full-length coding region of Bvh was 2190 bp, which encodes a 729 amino acid (aa) protein containing nine consensus regions of the DEAD box protein family. Bvh is expressed only in the ovary and testis of adult cattle. Two splice variants were identified and termed Bvh-V4 (2112 bp and 703 aa) and Bvh-V45 (2040 bp and 679 aa). In male cattle, full-length Bvh (Bvh-FL), Bvh-V4 and Bvh-V45 are exclusively expressed in the testes in the ratio of 2.2:1.6:1, respectively. Real-time PCR revealed significantly reduced mRNA expression of Bvh-FL, Bvh-V4 and Bvh-V45 in testes of cattle-yak hybrids, with meiotic arrest compared with cattle and yaks with normal spermatogenesis (P < 0.01). The promoter methylation level of Bvh in the testes of cattle-yak hybrids was significantly greater than in cattle and yaks (P < 0.01). Conclusion In the present study, Bvh was isolated and characterized. These data suggest that Bvh functions in bovine spermatogenesis, and that transcription of the gene in testes were regulated by alternative splice and promoter methylation. PMID:23815438

  8. Life cycle stage and water depth affect flooding-induced adventitious root formation in the terrestrial species Solanum dulcamara

    PubMed Central

    Zhang, Qian; Visser, Eric J. W.; de Kroon, Hans; Huber, Heidrun

    2015-01-01

    Background and Aims Flooding can occur at any stage of the life cycle of a plant, but often adaptive responses of plants are only studied at a single developmental stage. It may be anticipated that juvenile plants may respond differently from mature plants, as the amount of stored resources may differ and morphological changes can be constrained. Moreover, different water depths may require different strategies to cope with the flooding stress, the expression of which may also depend on developmental stage. This study investigated whether flooding-induced adventitious root formation and plant growth were affected by flooding depth in Solanum dulcamara plants at different developmental stages. Methods Juvenile plants without pre-formed adventitious root primordia and mature plants with primordia were subjected to shallow flooding or deep flooding for 5 weeks. Plant growth and the timing of adventitious root formation were monitored during the flooding treatments. Key Results Adventitious root formation in response to shallow flooding was significantly constrained in juvenile S. dulcamara plants compared with mature plants, and was delayed by deep flooding compared with shallow flooding. Complete submergence suppressed adventitious root formation until up to 2 weeks after shoots restored contact with the atmosphere. Independent of developmental stage, a strong positive correlation was found between adventitious root formation and total biomass accumulation during shallow flooding. Conclusions The potential to deploy an escape strategy (i.e. adventitious root formation) may change throughout a plant’s life cycle, and is largely dependent on flooding depth. Adaptive responses at a given stage of the life cycle thus do not necessarily predict how the plant responds to flooding in another growth stage. As variation in adventitious root formation also correlates with finally attained biomass, this variation may form the basis for variation in resistance to shallow

  9. Quantitative Analysis of Adventitious Root Growth Phenotypes in Carnation Stem Cuttings.

    PubMed

    Birlanga, Virginia; Villanova, Joan; Cano, Antonio; Cano, Emilio A; Acosta, Manuel; Pérez-Pérez, José Manuel

    2015-01-01

    Carnation is one of the most important species on the worldwide market of cut flowers. Commercial carnation cultivars are vegetatively propagated from terminal stem cuttings that undergo a rooting and acclimation process. For some of the new cultivars that are being developed by ornamental breeders, poor adventitious root (AR) formation limits its commercial scaling-up, due to a significant increase in the production costs. We have initiated a genetical-genomics approach to determine the molecular basis of the differences found between carnation cultivars during adventitious rooting. The detailed characterization of AR formation in several carnation cultivars differing in their rooting losses has been performed (i) during commercial production at a breeders' rooting station and (ii) on a defined media in a controlled environment. Our study reveals the phenotypic signatures that distinguishes the bad-rooting cultivars and provides the appropriate set-up for the molecular identification of the genes involved in AR development in this species.

  10. Quantitative Analysis of Adventitious Root Growth Phenotypes in Carnation Stem Cuttings

    PubMed Central

    Birlanga, Virginia; Villanova, Joan; Cano, Antonio; Cano, Emilio A.; Acosta, Manuel; Pérez-Pérez, José Manuel

    2015-01-01

    Carnation is one of the most important species on the worldwide market of cut flowers. Commercial carnation cultivars are vegetatively propagated from terminal stem cuttings that undergo a rooting and acclimation process. For some of the new cultivars that are being developed by ornamental breeders, poor adventitious root (AR) formation limits its commercial scaling-up, due to a significant increase in the production costs. We have initiated a genetical-genomics approach to determine the molecular basis of the differences found between carnation cultivars during adventitious rooting. The detailed characterization of AR formation in several carnation cultivars differing in their rooting losses has been performed (i) during commercial production at a breeders’ rooting station and (ii) on a defined media in a controlled environment. Our study reveals the phenotypic signatures that distinguishes the bad-rooting cultivars and provides the appropriate set-up for the molecular identification of the genes involved in AR development in this species. PMID:26230608

  11. The quiescent center and the stem cell niche in the adventitious roots of Arabidopsis thaliana

    PubMed Central

    Della Rovere, Federica; Fattorini, Laura; Ronzan, Marilena; Falasca, Giuseppina; Altamura, Maria Maddalena

    2016-01-01

    ABSTRACT Adventitious rooting is essential for the survival of numerous species from vascular cryptogams to monocots, and is required for successful micropropagation. The tissues involved in AR initiation may differ in planta and in in vitro systems. For example, in Arabidopsis thaliana, ARs originate from the hypocotyl pericycle in planta and the stem endodermis in in vitro cultured thin cell layers. The formation of adventitious roots (ARs) depends on numerous factors, among which the hormones, auxin, in particular. In both primary and lateral roots, growth depends on a functional stem cell niche in the apex, maintained by an active quiescent center (QC), and involving the expression of genes controlled by auxin and cytokinin. This review summarizes current knowledge about auxin and cytokinin control on genes involved in the definition and maintenance of QC, and stem cell niche, in the apex of Arabidopsis ARs in planta and in longitudinal thin cell layers. PMID:27089118

  12. Quantifying Adventitious Error in a Covariance Structure as a Random Effect.

    PubMed

    Wu, Hao; Browne, Michael W

    2015-09-01

    We present an approach to quantifying errors in covariance structures in which adventitious error, identified as the process underlying the discrepancy between the population and the structured model, is explicitly modeled as a random effect with a distribution, and the dispersion parameter of this distribution to be estimated gives a measure of misspecification. Analytical properties of the resultant procedure are investigated and the measure of misspecification is found to be related to the root mean square error of approximation. An algorithm is developed for numerical implementation of the procedure. The consistency and asymptotic sampling distributions of the estimators are established under a new asymptotic paradigm and an assumption weaker than the standard Pitman drift assumption. Simulations validate the asymptotic sampling distributions and demonstrate the importance of accounting for the variations in the parameter estimates due to adventitious error. Two examples are also given as illustrations.

  13. Influences of polar auxin transport on polarity of adventitious bud formation in hybrid populas

    SciTech Connect

    Kim, Myung Won ); Hackett, W. )

    1989-04-01

    The role of auxin and cytokinin distribution of polar regeneration of adventitious bud has been investigated. Explants from leaf midvein were labelled with {sup 14}C-NAA and {sup 14}C-BA and the radioactivity in proximal, mid, and distal portions was counted after 24h and 48h. Explants showing polar regeneration of buds on the proximal end showed a clear polar distribution of {sup 14}CNAA. Auxin transport inhibitors (NPA, TIBA) eliminated polar distribution of auxin and reduced polarity of bud formation and the total number of buds formed, but did not reduce callus formation. Increased concentration of Ca(NO{sub 3}){sub 2} decreased polarity of bud formation and increased the number of buds formed but did not affect the distribution of auxin of cytokinin. Some factor in addition to polar distribution of auxin or cytokinin-auxin ratio appears to influence the polarity of adventitious bud formation.

  14. The quiescent center and the stem cell niche in the adventitious roots of Arabidopsis thaliana.

    PubMed

    Rovere, Federica Della; Fattorini, Laura; Ronzan, Marilena; Falasca, Giuseppina; Altamura, Maria Maddalena

    2016-05-01

    Adventitious rooting is essential for the survival of numerous species from vascular cryptogams to monocots, and is required for successful micropropagation. The tissues involved in AR initiation may differ in planta and in in vitro systems. For example, in Arabidopsis thaliana, ARs originate from the hypocotyl pericycle in planta and the stem endodermis in in vitro cultured thin cell layers. The formation of adventitious roots (ARs) depends on numerous factors, among which the hormones, auxin, in particular. In both primary and lateral roots, growth depends on a functional stem cell niche in the apex, maintained by an active quiescent center (QC), and involving the expression of genes controlled by auxin and cytokinin. This review summarizes current knowledge about auxin and cytokinin control on genes involved in the definition and maintenance of QC, and stem cell niche, in the apex of Arabidopsis ARs in planta and in longitudinal thin cell layers. PMID:27089118

  15. Cystic adventitial disease of the popliteal artery: an infrequent cause of intermittent claudication

    PubMed Central

    Kauffman, Paulo; Kuzniec, Sergio; Sacilotto, Roberto; Teivelis, Marcelo Passos; Wolosker, Nelson; Tachibana, Adriano

    2014-01-01

    Intermittent claudication is frequently associated with atherosclerotic disease, but differential diagnosis must be sought in patients with no traditional risk factors. Cystic adventitial disease, of unknown etiology, most frequently affects the popliteal artery, and occasionally presents as intermittent claudication. We report a case of this disease and the surgical treatment, and discuss some aspects related to etiopathogenesis, diagnosis and treatment of this condition. PMID:25167336

  16. Gibberellins inhibit adventitious rooting in hybrid aspen and Arabidopsis by affecting auxin transport.

    PubMed

    Mauriat, Mélanie; Petterle, Anna; Bellini, Catherine; Moritz, Thomas

    2014-05-01

    Knowledge of processes involved in adventitious rooting is important to improve both fundamental understanding of plant physiology and the propagation of numerous plants. Hybrid aspen (Populus tremula × tremuloïdes) plants overexpressing a key gibberellin (GA) biosynthesis gene (AtGA20ox1) grow rapidly but have poor rooting efficiency, which restricts their clonal propagation. Therefore, we investigated the molecular basis of adventitious rooting in Populus and the model plant Arabidopsis. The production of adventitious roots (ARs) in tree cuttings is initiated from the basal stem region, and involves the interplay of several endogenous and exogenous factors. The roles of several hormones in this process have been characterized, but the effects of GAs have not been fully investigated. Here, we show that a GA treatment negatively affects the numbers of ARs produced by wild-type hybrid aspen cuttings. Furthermore, both hybrid aspen plants and intact Arabidopsis seedlings overexpressing AtGA20ox1, PttGID1.1 or PttGID1.3 genes (with a 35S promoter) produce few ARs, although ARs develop from the basal stem region of hybrid aspen and the hypocotyl of Arabidopsis. In Arabidopsis, auxin and strigolactones are known to affect AR formation. Our data show that the inhibitory effect of GA treatment on adventitious rooting is not mediated by perturbation of the auxin signalling pathway, or of the strigolactone biosynthetic and signalling pathways. Instead, GAs appear to act by perturbing polar auxin transport, in particular auxin efflux in hybrid aspen, and both efflux and influx in Arabidopsis.

  17. The role of strigolactones in photomorphogenesis of pea is limited to adventitious rooting.

    PubMed

    Urquhart, Shelley; Foo, Eloise; Reid, James B

    2015-03-01

    The recently discovered group of plant hormones, the strigolactones, have been implicated in regulating photomorphogenesis. We examined this extensively in our strigolactone synthesis and response mutants and could find no evidence to support a major role for strigolactone signaling in classic seedling photomorphogenesis (e.g. elongation and leaf expansion) in pea (Pisum sativum), consistent with two recent independent reports in Arabidopsis. However, we did find a novel effect of strigolactones on adventitious rooting in darkness. Strigolactone-deficient mutants, Psccd8 and Psccd7, produced significantly fewer adventitious roots than comparable wild-type seedlings when grown in the dark, but not when grown in the light. This observation in dark-grown plants did not appear to be due to indirect effects of other factors (e.g. humidity) as the constitutively de-etiolated mutant, lip1, also displayed reduced rooting in the dark. This role for strigolactones did not involve the MAX2 F-Box strigolactone response pathway as Psmax2 f-box mutants did not show a reduction in adventitious rooting in the dark compared with wild-type plants. The auxin-deficient mutant bushy also reduced adventitious rooting in the dark, as did decapitation of wild-type plants. Rooting was restored by the application of indole-3-acetic acid (IAA) to decapitated plants, suggesting a role for auxin in the rooting response. However, auxin measurements showed no accumulation of IAA in the epicotyls of wild-type plants compared with the strigolactone synthesis mutant Psccd8, suggesting that changes in the gross auxin level in the epicotyl are not mediating this response to strigolactone deficiency.

  18. Nitric oxide is involved in hemin-induced cucumber adventitious rooting process.

    PubMed

    Xuan, Wei; Xu, Sheng; Li, Meiyue; Han, Bin; Zhang, Bo; Zhang, Jing; Lin, Yuting; Huang, Jingjing; Shen, Wenbiao; Cui, Jin

    2012-07-15

    Hemin, a heme oxygenase-1 (HO-1) inducer, was shown to exert numerous beneficial physiological functions in animals. Our previous study suggests that HO-1/carbon monoxide (CO) acts as a novel downstream signal system in the auxin-induced adventitious rooting. The objective of this study was to test whether nitric oxide (NO) is involved in hemin-induced cucumber adventitious rooting. Applications of hemin or CO aqueous solution to auxin-depleted cucumber explant induced up-regulation of cucumber HO-1 transcripts (CsHO1), NO production, and thereafter adventitious root formation, and some above responses were blocked by the combination treatment with two nitric oxide synthase (NOS)-like enzyme inhibitors N(G)-nitro-L-arginine methylester hydrochloride and N(G)-nitro-L-arginine, a HO-1 specific inhibitor zinc protoporphyrin IX, and a specific NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt. However, these blocking responses were not observed using tungstate, an inhibitor of nitrate reductase, another NO producing enzyme in plants. Furthermore, the guanylate cyclase inhibitors 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalin-1-one and 6-anilino-5,8-quinolinedione reduced root development induced by hemin, whereas the cell-permeable cyclic guanosine monophosphate (cGMP) derivative 8-Br-cGMP reversed this effect. Together, our results indicated that at least in our experimental conditions, NO might operate downstream of hemin promoting adventitious root formation probably in a cGMP-dependent manner. PMID:22579358

  19. The role of strigolactones in photomorphogenesis of pea is limited to adventitious rooting.

    PubMed

    Urquhart, Shelley; Foo, Eloise; Reid, James B

    2015-03-01

    The recently discovered group of plant hormones, the strigolactones, have been implicated in regulating photomorphogenesis. We examined this extensively in our strigolactone synthesis and response mutants and could find no evidence to support a major role for strigolactone signaling in classic seedling photomorphogenesis (e.g. elongation and leaf expansion) in pea (Pisum sativum), consistent with two recent independent reports in Arabidopsis. However, we did find a novel effect of strigolactones on adventitious rooting in darkness. Strigolactone-deficient mutants, Psccd8 and Psccd7, produced significantly fewer adventitious roots than comparable wild-type seedlings when grown in the dark, but not when grown in the light. This observation in dark-grown plants did not appear to be due to indirect effects of other factors (e.g. humidity) as the constitutively de-etiolated mutant, lip1, also displayed reduced rooting in the dark. This role for strigolactones did not involve the MAX2 F-Box strigolactone response pathway as Psmax2 f-box mutants did not show a reduction in adventitious rooting in the dark compared with wild-type plants. The auxin-deficient mutant bushy also reduced adventitious rooting in the dark, as did decapitation of wild-type plants. Rooting was restored by the application of indole-3-acetic acid (IAA) to decapitated plants, suggesting a role for auxin in the rooting response. However, auxin measurements showed no accumulation of IAA in the epicotyls of wild-type plants compared with the strigolactone synthesis mutant Psccd8, suggesting that changes in the gross auxin level in the epicotyl are not mediating this response to strigolactone deficiency. PMID:24962787

  20. Detecting and quantifying the adventitious presence of transgenic seeds in safflower, Carthamus tinctorius L.

    PubMed

    Christianson, Jed; McPherson, Marc; Topinka, Deborah; Hall, Linda; Good, Allen G

    2008-07-23

    Safflower ( Carthamus tinctorius L.) is currently being developed as a platform for the production of novel proteins. Methods for detecting and quantifying transgenic safflower are needed to ensure seed quality and to monitor for its adventitious presence. We developed and compared three methods of assaying for transgenic safflower presence in conventional seedlots: field bioassays, enzyme-linked immunosorbent assays (ELISA), and quantitative polymerase chain reaction (Q-PCR). Limits for reliable quantification for both ELISA and Q-PCR are approximately 0.1%, although levels at least as low as 0.02% can be detected by Q-PCR. Levels of quantification for the field bioassay are limited only by space and resources available. Multiple sampling methods to detect and quantify transgenic safflower presence at levels lower than 0.1% were used on field collected samples from a pollen outcrossing experiment to quantify the adventitious presence of transgenic safflower. Taking into account the potential utility and relative advantages or disadvantages of each detection method, it is recommended that the initial testing for the adventitious presence of transgenic seed be carried out using an antibody-based test if available and that Q-PCR-based assays to quantify transgenic proportion be used when it is necessary to identify specific transgenic constructs or if antibody-based assays are not readily available.

  1. Large Scale Culture of Ginseng Adventitious Roots for Production of Ginsenosides

    NASA Astrophysics Data System (ADS)

    Paek, Kee-Yoeup; Murthy, Hosakatte Niranjana; Hahn, Eun-Joo; Zhong, Jian-Jiang

    Ginseng (Panax ginseng C. A. Meyer) is one of the most famous oriental medicinal plants used as crude drugs in Asian countries, and now it is being used worldwide for preventive and therapeutic purposes. Among diverse constituents of ginseng, saponins (ginsenosides) have been found to be major components responsible for their biological and pharmacological actions. On the other hand, difficulties in the supply of pure ginsenosides in quantity prevent the development of ginseng for clinical medicines. Cultivation of ginseng in fields takes a long time, generally 5-7 years, and needs extensive effort regarding quality control since growth is susceptible to many environmental factors including soil, shade, climate, pathogens and pests. To solve the problems, cell and tissue cultures have been widely explored for more rapid and efficient production of ginseng biomass and ginsenosides. Recently, cell and adventitious root cultures of P. ginseng have been established in large scale bioreactors with a view to commercial application. Various physiological and engineering parameters affecting the biomass production and ginsenoside accumulation have been investigated. Advances in adventitious root cultures including factors for process scale-up are reviewed in this chapter. In addition, biosafety analyses of ginseng adventitious roots are also discussed for real application.

  2. Proper gibberellin localization in vascular tissue is required to regulate adventitious root development in tobacco.

    PubMed

    Niu, Shihui; Li, Zhexin; Yuan, Huwei; Fang, Pan; Chen, Xiaoyang; Li, Wei

    2013-08-01

    Bioactive gibberellins (GAs) are involved in many developmental aspects of the life cycle of plants, acting either directly or through interaction with other hormones. Accumulating evidence suggests that GAs have an important effect on root growth; however, there is currently little information on the specific regulatory mechanism of GAs during adventitious root development. A study was conducted on tobacco (Nicotiana tabacum) plants for altered rates of biosynthesis, catabolism, and GA signalling constitutively or in specific tissues using a transgenic approach. In the present study, PtGA20ox, PtGA2ox1, and PtGAI were overexpressed under the control of the 35S promoter, vascular cambium-specific promoter (LMX5), or root meristem-specific promoter (TobRB7), respectively. Evidence is provided that the precise localization of bioactive GA in the stem but not in the roots is required to regulate adventitious root development in tobacco. High levels of GA negatively regulate the early initiation step of root formation through interactions with auxin, while a proper and mobile GA signal is required for the emergence and subsequent long-term elongation of established primordia. The results demonstrated that GAs have an inhibitory effect on adventitious root formation but a stimulatory effect on root elongation. PMID:23918971

  3. vasa-related genes and their expression in stem cells of colonial parasitic rhizocephalan barnacle Polyascus polygenea (Arthropoda: Crustacea: Cirripedia: Rhizocephala).

    PubMed

    Shukalyuk, Andrey I; Golovnina, Kseniya A; Baiborodin, Sergei I; Gunbin, Konstantin V; Blinov, Alexander G; Isaeva, Valeria V

    2007-02-01

    vasa (vas)-related genes are members of the DEAD-box protein family and are expressed in the germ cells of many Metazoa. We cloned vasa-related genes (PpVLG, CpVLG) and other DEAD-box family related genes (PpDRH1, PpDRH2, CpDRH, AtDRHr) from the colonial parasitic rhizocephalan barnacle Polyascus polygenea, the non-colonial Clistosaccus paguri (Crustacea: Cirripedia: Rhizocephala), and the parasitic isopodan Athelgis takanoshimensis (Crustacea: Isopoda). The colonial Polyascus polygenea, a parasite of the coastal crabs Hemigrapsus sanguineus and Hemigrapsus longitarsis was used as a model object for further detailed investigations. Phylogenetic analysis suggested that PpVLG and CpVLG are closely related to vasa-like genes of other Arthropoda. The rest of the studied genes form their own separate branch on the phylogenetic tree and have a common ancestry with the p68 and PL10 subfamilies. We suppose this group may be a new subfamily of the DEAD-box RNA helicases that is specific for parasitic Crustacea. We found PpVLG and PpDRH1 expression products in stem cells from stolons and buds of internae, during asexual reproduction of colonial P. polygenea, and in germ cells from sexually reproducing externae, including male spermatogenic cells and female oogenic cells.

  4. Transcriptome analysis of indole-3-butyric acid-induced adventitious root formation in nodal cuttings of Camellia sinensis (L.).

    PubMed

    Wei, Kang; Wang, Li-Yuan; Wu, Li-Yun; Zhang, Cheng-Cai; Li, Hai-Lin; Tan, Li-Qiang; Cao, Hong-Li; Cheng, Hao

    2014-01-01

    Tea (Camellia sinensis L.) is a popular world beverage, and propagation of tea plants chiefly depends on the formation of adventitious roots in cuttings. To better understand potential mechanisms involved in adventitious root formation, we performed transcriptome analysis of single nodal cuttings of C. sinensis treated with or without indole-3-butyric acid (IBA) using the Illumina sequencing method. Totally 42.5 million RNA-Seq reads were obtained and these were assembled into 59,931 unigenes, with an average length of 732 bp and an N50 of 1292 bp. In addition, 1091 differentially expressed unigenes were identified in the tea cuttings treated with IBA compared to controls, including 656 up- and 435 down-regulated genes. Further real time RT-PCR analysis confirmed RNA-Seq data. Functional annotation analysis showed that many genes were involved in plant hormone signal transduction, secondary metabolism, cell wall organization and glutathione metabolism, indicating potential contributions to adventitious rooting. Our study presents a global view of transcriptome profiles of tea cuttings in response to IBA treatment and provides new insights into the fundamental mechanisms associated with auxin-induced adventitious rooting. Our data will be a valuable resource for genomic research about adventitious root formation in tea cuttings, which can be used to improve rooting for difficult-to-root varieties.

  5. Transcriptome Analysis of Indole-3-Butyric Acid-Induced Adventitious Root Formation in Nodal Cuttings of Camellia sinensis (L.)

    PubMed Central

    Wei, Kang; Wang, Li-Yuan; Wu, Li-Yun; Zhang, Cheng-Cai; Li, Hai-Lin; Tan, Li-Qiang; Cao, Hong-Li; Cheng, Hao

    2014-01-01

    Tea (Camellia sinensis L.) is a popular world beverage, and propagation of tea plants chiefly depends on the formation of adventitious roots in cuttings. To better understand potential mechanisms involved in adventitious root formation, we performed transcriptome analysis of single nodal cuttings of C. sinensis treated with or without indole-3-butyric acid (IBA) using the Illumina sequencing method. Totally 42.5 million RNA-Seq reads were obtained and these were assembled into 59,931 unigenes, with an average length of 732 bp and an N50 of 1292 bp. In addition, 1091 differentially expressed unigenes were identified in the tea cuttings treated with IBA compared to controls, including 656 up- and 435 down-regulated genes. Further real time RT-PCR analysis confirmed RNA-Seq data. Functional annotation analysis showed that many genes were involved in plant hormone signal transduction, secondary metabolism, cell wall organization and glutathione metabolism, indicating potential contributions to adventitious rooting. Our study presents a global view of transcriptome profiles of tea cuttings in response to IBA treatment and provides new insights into the fundamental mechanisms associated with auxin-induced adventitious rooting. Our data will be a valuable resource for genomic research about adventitious root formation in tea cuttings, which can be used to improve rooting for difficult-to-root varieties. PMID:25216187

  6. Transcriptome analysis of indole-3-butyric acid-induced adventitious root formation in nodal cuttings of Camellia sinensis (L.).

    PubMed

    Wei, Kang; Wang, Li-Yuan; Wu, Li-Yun; Zhang, Cheng-Cai; Li, Hai-Lin; Tan, Li-Qiang; Cao, Hong-Li; Cheng, Hao

    2014-01-01

    Tea (Camellia sinensis L.) is a popular world beverage, and propagation of tea plants chiefly depends on the formation of adventitious roots in cuttings. To better understand potential mechanisms involved in adventitious root formation, we performed transcriptome analysis of single nodal cuttings of C. sinensis treated with or without indole-3-butyric acid (IBA) using the Illumina sequencing method. Totally 42.5 million RNA-Seq reads were obtained and these were assembled into 59,931 unigenes, with an average length of 732 bp and an N50 of 1292 bp. In addition, 1091 differentially expressed unigenes were identified in the tea cuttings treated with IBA compared to controls, including 656 up- and 435 down-regulated genes. Further real time RT-PCR analysis confirmed RNA-Seq data. Functional annotation analysis showed that many genes were involved in plant hormone signal transduction, secondary metabolism, cell wall organization and glutathione metabolism, indicating potential contributions to adventitious rooting. Our study presents a global view of transcriptome profiles of tea cuttings in response to IBA treatment and provides new insights into the fundamental mechanisms associated with auxin-induced adventitious rooting. Our data will be a valuable resource for genomic research about adventitious root formation in tea cuttings, which can be used to improve rooting for difficult-to-root varieties. PMID:25216187

  7. Growth and anatomical parameters of adventitious roots formed on mung bean hypocotyls are correlated with galactoglucomannan oligosaccharides structure.

    PubMed

    Kollárová, K; Zelko, I; Henselová, M; Capek, P; Lišková, D

    2012-01-01

    The effect of galactoglucomannan oligosaccharides (GGMOs) compared with chemically modified oligosaccharides, GGMOs-g (with reduced number of D-galactose side chains) and GGMOs-r (with reduced reducing ends) on mung bean (Vigna radiata (L.) Wilczek) adventitious roots formation, elongation, and anatomical structure have been studied. All types of oligosaccharides influenced adventitious root formation in the same way: stimulation in the absence of exogenous auxin and inhibition in the presence of exogenous auxin. Both reactions are probably related with the presence/content of endogenous auxin in plant cuttings. However, the adventitious root length was inhibited by GGMOs both in the absence as well as in the presence of auxin (IBA or NAA), while GGMOs-g inhibition was significantly weaker compared with GGMOs. GGMOs-r were without significant difference on both processes, compared with GGMOs. GGMOs affected not only the adventitious root length but also their anatomy in dependence on the combination with certain type of auxin. The oligosaccharides influenced cortical cells division, which was reflected in the cortex area and in the root diameter. All processes followed were dependent on oligosaccharides chemical structure. The results suggest also that GGM-derived oligosaccharides may play an important role in adventitious roots elongation but not in their formation. PMID:22666154

  8. Adventitial Cells and Perictyes Support Chondrogenesis Through Different Mechanisms in 3-Dimensional Cultures With or Without Nanoscaffolds.

    PubMed

    Zhang, Shu; Ba, Kai; Wu, Ling; Lee, Siyong; Peault, Bruno; Petrigliano, Frank A; McAllister, David R; Adams, John S; Evseenko, Denis; Lin, Yunfeng

    2015-10-01

    In previous studies, mesenchymal stromal cells (MSCs) derived from bone marrow and fat tissues were shown to increase proliferation and matrix production of chondrocytes (CH) in co-culture. The aim of this study was to investigate the roles of pericytes (CD31(neg)CD45(neg)CD146+CD34(neg)) and adventitial cells (CD31(neg)CD45(neg)CD146(neg)CD34+) sub-populations of MSCs in supporting proliferation and matrix deposition of CH. The MSCs were derived from synovial membrane and attaching fat tissue. Then, the pericytes and adventitial cells were sorted from total MSCs and co-cultured with articular CH respectively. In pellet co-culture model, the pericytes showed more prominent effects on glycosaminoglycans (GAGs) production and Collagen II synthesis than the adventitial cells which had stronger effects on promoting CH proliferation. In addition, quantitative polymerase chain reaction (qPCR) was performed to examine the expression of a group of secreted growth factors and co-culture performed on electrospun scaffolds based on Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB), to verify the trophic effects of different MSC sub-populations in 3-Dimensional (3D) environment. In conclusion, it was found that the pericytes and adventitial cells support CH in different ways; the adventitial cells more supporting the proliferation of CH, while pericytes are better in stimulating GAGs and collagen production of CH. PMID:26502642

  9. Germ plasm localisation of the HELICc of Vasa in Drosophila: analysis of domain sufficiency and amino acids critical for localisation

    PubMed Central

    Wang, Szu-Chieh; Hsu, Hao-Jen; Lin, Gee-way; Wang, Ting-Fang; Chang, Chun-che; Lin, Ming-Der

    2015-01-01

    Formation of the germ plasm drives germline specification in Drosophila and some other insects such as aphids. Identification of the DEAD-box protein Vasa (Vas) as a conserved germline marker in flies and aphids suggests that they share common components for assembling the germ plasm. However, to which extent the assembly order is conserved and the correlation between functions and sequences of Vas remain unclear. Ectopic expression of the pea aphid Vas (ApVas1) in Drosophila did not drive its localisation to the germ plasm, but ApVas1 with a replaced C-terminal domain (HELICc) of Drosophila Vas (DmVas) became germ-plasm restricted. We found that HELICc itself, through the interaction with Oskar (Osk), was sufficient for germ-plasm localisation. Similarly, HELICc of the grasshopper Vas could be recruited to the germ plasm in Drosophila. Nonetheless, germ-plasm localisation was not seen in the Drosophila oocytes expressing HELICcs of Vas orthologues from aphids, crickets, and mice. We further identified that glutamine (Gln) 527 within HELICc of DmVas was critical for localisation, and its corresponding residue could also be detected in grasshopper Vas yet missing in the other three species. This suggests that Gln527 is a direct target of Osk or critical to the maintenance of HELICc conformation. PMID:26419889

  10. Germ plasm localisation of the HELICc of Vasa in Drosophila: analysis of domain sufficiency and amino acids critical for localisation

    NASA Astrophysics Data System (ADS)

    Wang, Szu-Chieh; Hsu, Hao-Jen; Lin, Gee-Way; Wang, Ting-Fang; Chang, Chun-Che; Lin, Ming-Der

    2015-09-01

    Formation of the germ plasm drives germline specification in Drosophila and some other insects such as aphids. Identification of the DEAD-box protein Vasa (Vas) as a conserved germline marker in flies and aphids suggests that they share common components for assembling the germ plasm. However, to which extent the assembly order is conserved and the correlation between functions and sequences of Vas remain unclear. Ectopic expression of the pea aphid Vas (ApVas1) in Drosophila did not drive its localisation to the germ plasm, but ApVas1 with a replaced C-terminal domain (HELICc) of Drosophila Vas (DmVas) became germ-plasm restricted. We found that HELICc itself, through the interaction with Oskar (Osk), was sufficient for germ-plasm localisation. Similarly, HELICc of the grasshopper Vas could be recruited to the germ plasm in Drosophila. Nonetheless, germ-plasm localisation was not seen in the Drosophila oocytes expressing HELICcs of Vas orthologues from aphids, crickets, and mice. We further identified that glutamine (Gln) 527 within HELICc of DmVas was critical for localisation, and its corresponding residue could also be detected in grasshopper Vas yet missing in the other three species. This suggests that Gln527 is a direct target of Osk or critical to the maintenance of HELICc conformation.

  11. Germ plasm localisation of the HELICc of Vasa in Drosophila: analysis of domain sufficiency and amino acids critical for localisation.

    PubMed

    Wang, Szu-Chieh; Hsu, Hao-Jen; Lin, Gee-way; Wang, Ting-Fang; Chang, Chun-che; Lin, Ming-Der

    2015-09-30

    Formation of the germ plasm drives germline specification in Drosophila and some other insects such as aphids. Identification of the DEAD-box protein Vasa (Vas) as a conserved germline marker in flies and aphids suggests that they share common components for assembling the germ plasm. However, to which extent the assembly order is conserved and the correlation between functions and sequences of Vas remain unclear. Ectopic expression of the pea aphid Vas (ApVas1) in Drosophila did not drive its localisation to the germ plasm, but ApVas1 with a replaced C-terminal domain (HELICc) of Drosophila Vas (DmVas) became germ-plasm restricted. We found that HELICc itself, through the interaction with Oskar (Osk), was sufficient for germ-plasm localisation. Similarly, HELICc of the grasshopper Vas could be recruited to the germ plasm in Drosophila. Nonetheless, germ-plasm localisation was not seen in the Drosophila oocytes expressing HELICcs of Vas orthologues from aphids, crickets, and mice. We further identified that glutamine (Gln) 527 within HELICc of DmVas was critical for localisation, and its corresponding residue could also be detected in grasshopper Vas yet missing in the other three species. This suggests that Gln527 is a direct target of Osk or critical to the maintenance of HELICc conformation.

  12. Bone morphogenetic protein 4 and retinoic acid trigger bovine VASA homolog expression in differentiating bovine induced pluripotent stem cells.

    PubMed

    Malaver-Ortega, Luis F; Sumer, Huseyin; Jain, Kanika; Verma, Paul J

    2016-02-01

    Primordial germ cells (PGCs) are the earliest identifiable and completely committed progenitors of female and male gametes. They are obvious targets for genome editing because they assure the transmission of desirable or introduced traits to future generations. PGCs are established at the earliest stages of embryo development and are difficult to propagate in vitro--two characteristics that pose a problem for their practical application. One alternative method to enrich for PGCs in vitro is to differentiate them from pluripotent stem cells derived from adult tissues. Here, we establish a reporter system for germ cell identification in bovine pluripotent stem cells based on green fluorescent protein expression driven by the minimal essential promoter of the bovine Vasa homolog (BVH) gene, whose regulatory elements were identified by orthologous modelling of regulatory units. We then evaluated the potential of bovine induced pluripotent stem cell (biPSC) lines carrying the reporter construct to differentiate toward the germ cell lineage. Our results showed that biPSCs undergo differentiation as embryoid bodies, and a fraction of the differentiating cells expressed BVH. The rate of differentiation towards BVH-positive cells increased up to tenfold in the presence of bone morphogenetic protein 4 or retinoic acid. Finally, we determined that the expression of key PGC genes, such as BVH or SOX2, can be modified by pre-differentiation cell culture conditions, although this increase is not necessarily mirrored by an increase in the rate of differentiation.

  13. Aquatic adventitious root development in partially and completely submerged wetland plants Cotula coronopifolia and Meionectes brownii

    PubMed Central

    Rich, Sarah Meghan; Ludwig, Martha; Colmer, Timothy David

    2012-01-01

    Background and Aims A common response of wetland plants to flooding is the formation of aquatic adventitious roots. Observations of aquatic root growth are widespread; however, controlled studies of aquatic roots of terrestrial herbaceous species are scarce. Submergence tolerance and aquatic root growth and physiology were evaluated in two herbaceous, perennial wetland species Cotula coronopifolia and Meionectes brownii. Methods Plants were raised in large pots with ‘sediment’ roots in nutrient solution and then placed into individual tanks and shoots were left in air or submerged (completely or partially). The effects on growth of aquatic root removal, and of light availability to submerged plant organs, were evaluated. Responses of aquatic root porosity, chlorophyll and underwater photosynthesis, were studied. Key Results Both species tolerated 4 weeks of complete or partial submergence. Extensive, photosynthetically active, aquatic adventitious roots grew from submerged stems and contributed up to 90 % of the total root dry mass. When aquatic roots were pruned, completely submerged plants grew less and had lower stem and leaf chlorophyll a, as compared with controls with intact roots. Roots exposed to the lowest PAR (daily mean 4·7 ± 2·4 µmol m−2 s−1) under water contained less chlorophyll, but there was no difference in aquatic root biomass after 4 weeks, regardless of light availability in the water column (high PAR was available to all emergent shoots). Conclusions Both M. brownii and C. coronopifolia responded to submergence with growth of aquatic adventitious roots, which essentially replaced the existing sediment root system. These aquatic roots contained chlorophyll and were photosynthetically active. Removal of aquatic roots had negative effects on plant growth during partial and complete submergence. PMID:22419759

  14. Development of a candidate reference material for adventitious virus detection in vaccine and biologicals manufacturing by deep sequencing

    PubMed Central

    Mee, Edward T.; Preston, Mark D.; Minor, Philip D.; Schepelmann, Silke; Huang, Xuening; Nguyen, Jenny; Wall, David; Hargrove, Stacey; Fu, Thomas; Xu, George; Li, Li; Cote, Colette; Delwart, Eric; Li, Linlin; Hewlett, Indira; Simonyan, Vahan; Ragupathy, Viswanath; Alin, Voskanian-Kordi; Mermod, Nicolas; Hill, Christiane; Ottenwälder, Birgit; Richter, Daniel C.; Tehrani, Arman; Jacqueline, Weber-Lehmann; Cassart, Jean-Pol; Letellier, Carine; Vandeputte, Olivier; Ruelle, Jean-Louis; Deyati, Avisek; La Neve, Fabio; Modena, Chiara; Mee, Edward; Schepelmann, Silke; Preston, Mark; Minor, Philip; Eloit, Marc; Muth, Erika; Lamamy, Arnaud; Jagorel, Florence; Cheval, Justine; Anscombe, Catherine; Misra, Raju; Wooldridge, David; Gharbia, Saheer; Rose, Graham; Ng, Siemon H.S.; Charlebois, Robert L.; Gisonni-Lex, Lucy; Mallet, Laurent; Dorange, Fabien; Chiu, Charles; Naccache, Samia; Kellam, Paul; van der Hoek, Lia; Cotten, Matt; Mitchell, Christine; Baier, Brian S.; Sun, Wenping; Malicki, Heather D.

    2016-01-01

    Background Unbiased deep sequencing offers the potential for improved adventitious virus screening in vaccines and biotherapeutics. Successful implementation of such assays will require appropriate control materials to confirm assay performance and sensitivity. Methods A common reference material containing 25 target viruses was produced and 16 laboratories were invited to process it using their preferred adventitious virus detection assay. Results Fifteen laboratories returned results, obtained using a wide range of wet-lab and informatics methods. Six of 25 target viruses were detected by all laboratories, with the remaining viruses detected by 4–14 laboratories. Six non-target viruses were detected by three or more laboratories. Conclusion The study demonstrated that a wide range of methods are currently used for adventitious virus detection screening in biological products by deep sequencing and that they can yield significantly different results. This underscores the need for common reference materials to ensure satisfactory assay performance and enable comparisons between laboratories. PMID:26709640

  15. Panax ginseng Adventitious Root Suspension Culture: Protocol for Biomass Production and Analysis of Ginsenosides by High Pressure Liquid Chromatography.

    PubMed

    Murthy, Hosakatte Niranjana; Paek, Kee Yoeup

    2016-01-01

    Panax ginseng C.A. Meyer (Korean ginseng) is a popular herbal medicine. It has been used in Chinese and Oriental medicines since thousands of years. Ginseng products are generally used as a tonic and an adaptogen to resist the adverse influence of a wide range of physical, chemical and biological factors, and to restore homeostasis. Ginsenosides or ginseng saponins are the principal active ingredients of ginseng. Since ginseng cultivation process is very slow and needs specific environment for field cultivation, cell and tissue cultures are sought as alternatives for the production of ginseng biomass and bioactive compounds. In this chapter, we focus on methods of induction of adventitious roots from ginseng roots, establishment of adventitious root suspension cultures using bioreactors, procedures for processing of adventitious roots, and analysis of ginsenosides by high pressure liquid chromatography. PMID:27108314

  16. Panax ginseng Adventitious Root Suspension Culture: Protocol for Biomass Production and Analysis of Ginsenosides by High Pressure Liquid Chromatography.

    PubMed

    Murthy, Hosakatte Niranjana; Paek, Kee Yoeup

    2016-01-01

    Panax ginseng C.A. Meyer (Korean ginseng) is a popular herbal medicine. It has been used in Chinese and Oriental medicines since thousands of years. Ginseng products are generally used as a tonic and an adaptogen to resist the adverse influence of a wide range of physical, chemical and biological factors, and to restore homeostasis. Ginsenosides or ginseng saponins are the principal active ingredients of ginseng. Since ginseng cultivation process is very slow and needs specific environment for field cultivation, cell and tissue cultures are sought as alternatives for the production of ginseng biomass and bioactive compounds. In this chapter, we focus on methods of induction of adventitious roots from ginseng roots, establishment of adventitious root suspension cultures using bioreactors, procedures for processing of adventitious roots, and analysis of ginsenosides by high pressure liquid chromatography.

  17. A Case of Cystic Adventitial Degeneration of the Left Popliteal Artery Diagnosed by Intravascular Ultrasound

    PubMed Central

    Niizeki, Takeshi; Ishino, Mitsunori; Kitahara, Tatsuro; Yamauchi, So; Ikeno, Eiichiro; Kubota, Isao

    2016-01-01

    An 87-year-old male was admitted with intermittent claudication of the left calf. We performed lower extremity angiography, which revealed stenosis of the left popliteal artery. Intravascular ultrasound (IVUS) image correctly identified the cystic appearance of visualized extravascular hypodensity, causing extrinsic compression of the lumen. We diagnosed the condition as cystic adventitial degeneration (CAD) of the popliteal artery. We operated a resection of a cyst with the artery and replaced the autovein graft (saphenous vein). After surgery, the patient was free of symptoms. CAD is a rare disease; thus, our IVUS findings may provide unique diagnostic clues in patients with CAD. PMID:26949345

  18. Hydrogen peroxide is a second messenger in the salicylic acid-triggered adventitious rooting process in mung bean seedlings.

    PubMed

    Yang, Wei; Zhu, Changhua; Ma, Xiaoling; Li, Guijun; Gan, Lijun; Ng, Denny; Xia, Kai

    2013-01-01

    In plants, salicylic acid (SA) is a signaling molecule that regulates disease resistance responses, such as systemic acquired resistance (SAR) and hypertensive response (HR). SA has been implicated as participating in various biotic and abiotic stresses. This study was conducted to investigate the role of SA in adventitious root formation (ARF) in mung bean (Phaseolus radiatus L) hypocotyl cuttings. We observed that hypocotyl treatment with SA could significantly promote the adventitious root formation, and its effects were dose and time dependent. Explants treated with SA displayed a 130% increase in adventitious root number compared with control seedlings. The role of SA in mung bean hypocotyl ARF as well as its interaction with hydrogen peroxide (H2O2) were also elucidated. Pretreatment of mung bean explants with N, N'-dimethylthiourea (DMTU), a scavenger for H2O2, resulted in a significant reduction of SA-induced ARF. Diphenyleneiodonium (DPI), a specific inhibitor of membrane-linked NADPH oxidase, also inhibited the effect of adventitious rooting triggered by SA treatment. The determination of the endogenous H2O2 level indicated that the seedlings treated with SA could induce H2O2 accumulation compared with the control treatment. Our results revealed a distinctive role of SA in the promotion of adventitious rooting via the process of H2O2 accumulation. This conclusion was further supported by antioxidant enzyme activity assays. Based on these results, we conclude that the accumulation of free H2O2 might be a downstream event in response to SA-triggered adventitious root formation in mung bean seedlings.

  19. Adventitious match probability for autosomal profiles when primer binding site mutation is possible.

    PubMed

    Pope, Susan; Evett, Ian; Puch-Solis, Roberto

    2016-09-01

    This paper considers the situation where two DNA systems with differing primers have been used to produce DNA profiles for loading and searching of a DNA Database. With any profiling system there exists the possibility of a "primer binding site mutation" (PBSM). When such a mutation occurs at one of the loci in a profile, it has the effect that the associated allele is not visible in the profile. In the case where a person has two different alleles at a given locus (heterozygous) the effect of a PBSM would be that the profile would appear to be that of an individual with only one allele at that locus (homozygous). The paper investigates the potential for an adventitious match as a result of a PBSM when, for example, a crime profile and person profile that have originated from two different individuals are found to be the same as a result of a PBSM in one of the profiles. It is demonstrated, both by theory and using simulations, that the effect of PBSMs is to slightly decrease the adventitious match probability from what it would had the same DNA system been used. PMID:27420391

  20. A Methodology for Concomitant Isolation of Intimal and Adventitial Endothelial Cells from the Human Thoracic Aorta

    PubMed Central

    Leclercq, Anne; Veillat, Véronique; Loriot, Sandrine; Spuul, Pirjo; Madonna, Francesco; Roques, Xavier; Génot, Elisabeth

    2015-01-01

    Background Aortic diseases are diverse and involve a multiplicity of biological systems in the vascular wall. Aortic dissection, which is usually preceded by aortic aneurysm, is a leading cause of morbidity and mortality in modern societies. Although the endothelium is now known to play an important role in vascular diseases, its contribution to aneurysmal aortic lesions remains largely unknown. The aim of this study was to define a reliable methodology for the isolation of aortic intimal and adventitial endothelial cells in order to throw light on issues relevant to endothelial cell biology in aneurysmal diseases. Methodology/Principal Findings We set up protocols to isolate endothelial cells from both the intima and the adventitia of human aneurysmal aortic vessel segments. Throughout the procedure, analysis of cell morphology and endothelial markers allowed us to select an endothelial fraction which after two rounds of expansion yielded a population of >90% pure endothelial cells. These cells have the features and functionalities of freshly isolated cells and can be used for biochemical studies. The technique was successfully used for aortic vessel segments of 20 patients and 3 healthy donors. Conclusions/Significance This simple and highly reproducible method allows the simultaneous preparation of reasonably pure primary cultures of intimal and adventitial human endothelial cells, thus providing a reliable source for investigating their biology and involvement in both thoracic aneurysms and other aortic diseases. PMID:26599408

  1. Cystic adventitial disease of the common femoral vein: A case report.

    PubMed

    Yu, Jimei; Lu, Chaojun; Pan, Xiaohua; Li, Weihua

    2016-08-01

    Adventitial cystic disease (ACD) of the common femoral vein is a rare vascular disorder. It becomes more difficult to recognize preoperatively especially when the femoral vein is affected. We report the case of a 62-year-old female patient who presented with a one-month history of painless swelling in her right lower extremity. She had no specific past medical history and no history of trauma, and had a full coagulopathy profile that was negative for any hypercoagulable syndrome. On examination, her lower right leg was significantly swollen with a palpable mass in her right inguinal region. A computerized tomography (CT) with contrast was performed to provide more information and revealed an eccentric compression over the medial wall of the right common femoral vein. During surgical exploration, adventitial cystic mucinous disease was enucleated and the patient underwent femoral exploration, excision of the cysts and reconstruction of iliac femoral vein graft using an artificial blood vessel. The pathological examination confirmed the diagnosis. The patient continued to do well, and she had an unremarkable venous duplex evaluation at her 6-month follow-up. The presentation, investigation, treatment, and pathology of this condition are discussed with a literature review. PMID:27672549

  2. Cystic adventitial disease of the common femoral vein: A case report

    PubMed Central

    Yu, Jimei; Lu, Chaojun; Pan, Xiaohua; Li, Weihua

    2016-01-01

    Summary Adventitial cystic disease (ACD) of the common femoral vein is a rare vascular disorder. It becomes more difficult to recognize preoperatively especially when the femoral vein is affected. We report the case of a 62-year-old female patient who presented with a one-month history of painless swelling in her right lower extremity. She had no specific past medical history and no history of trauma, and had a full coagulopathy profile that was negative for any hypercoagulable syndrome. On examination, her lower right leg was significantly swollen with a palpable mass in her right inguinal region. A computerized tomography (CT) with contrast was performed to provide more information and revealed an eccentric compression over the medial wall of the right common femoral vein. During surgical exploration, adventitial cystic mucinous disease was enucleated and the patient underwent femoral exploration, excision of the cysts and reconstruction of iliac femoral vein graft using an artificial blood vessel. The pathological examination confirmed the diagnosis. The patient continued to do well, and she had an unremarkable venous duplex evaluation at her 6-month follow-up. The presentation, investigation, treatment, and pathology of this condition are discussed with a literature review. PMID:27672549

  3. Cystic adventitial disease of the popliteal artery: Two case reports and a review of the literature.

    PubMed

    Del Canto Peruyera, P; Vázquez, M J Vallina-Victorero; Velasco, M Botas; Álvarez, P Calvín; Salgado, A Álvarez; Álvarez, J Cerviño; Fernández, L J Álvarez

    2015-04-01

    Two cases of cystic adventitial disease treated at our institution over the last year are presented. They were middle-aged and apparently healthy patients, and the symptoms begin with a sudden onset of unilateral claudication. After performing a magnetic resonance angiography, a cystic formation attached to the adventitia of the popliteal artery was identified. Both patients were treated in the same manner, with resection of the affected arterial segment and vein bypass interposition. Both remain asymptomatic after one year of follow-up in one case and six months in the other. Cystic adventitial disease is a rare entity, which presents in patients without cardiovascular risk factors, so sometimes it takes long to reach a definitive diagnosis. Concerning the different treatment options, cyst excision together with the affected arterial segment seems to offer better mid- and long-term results when compared with other treatment options such as cyst aspiration or endovascular techniques, although there are no multicenter trials evidencing the superiority of one against the others.

  4. Cystic adventitial disease of the common femoral vein: A case report

    PubMed Central

    Yu, Jimei; Lu, Chaojun; Pan, Xiaohua; Li, Weihua

    2016-01-01

    Summary Adventitial cystic disease (ACD) of the common femoral vein is a rare vascular disorder. It becomes more difficult to recognize preoperatively especially when the femoral vein is affected. We report the case of a 62-year-old female patient who presented with a one-month history of painless swelling in her right lower extremity. She had no specific past medical history and no history of trauma, and had a full coagulopathy profile that was negative for any hypercoagulable syndrome. On examination, her lower right leg was significantly swollen with a palpable mass in her right inguinal region. A computerized tomography (CT) with contrast was performed to provide more information and revealed an eccentric compression over the medial wall of the right common femoral vein. During surgical exploration, adventitial cystic mucinous disease was enucleated and the patient underwent femoral exploration, excision of the cysts and reconstruction of iliac femoral vein graft using an artificial blood vessel. The pathological examination confirmed the diagnosis. The patient continued to do well, and she had an unremarkable venous duplex evaluation at her 6-month follow-up. The presentation, investigation, treatment, and pathology of this condition are discussed with a literature review.

  5. Efficacy of Vasa Avaleha and its granules on Tamaka Shwasa (bronchial asthma): Open-label randomized clinical study

    PubMed Central

    Paneliya, Ankit M.; Patgiri, Biswajyoti; Galib, R.; Prajapati, Pradeep Kumar

    2015-01-01

    Introduction: Bronchial asthma is one of the chronic inflammatory disorders of the respiratory tract causing a huge number of deaths annually. Increased industrialization and pollution are the exacerbating factors for this situation. In Ayurveda, this miserable condition is comparable with Tamaka Shwasa. Synthetic drugs provide instant symptomatic relief in cases of bronchial asthma but are known to develop certain adverse drug reactions. Considering this, the current suffering population is looking hopefully towards other systems of medicine such as Ayurveda for better relief. Ayurveda has a number of formulations to treat Tamaka Shwasa and is in practice with proven efficacy. Aims: To evaluate comparative clinical efficacy of Vasa Avaleha (VA) and its granules (GVA) in cases of Tamaka Shwasa. Materials and Methods: A total of 66 patients were registered and randomly grouped into A and B. Patients of Group A were treated with VA, while Group B with GVA at dose of 6 g twice a day with lukewarm water for the duration of 28 days. Follow-up was done after 14 days. The results were assessed in terms of clinical recovery, symptomatic relief, and pulmonary function improvement. Effect of the treatment was assessed based on subjective and objective parameters. Results: Significant improvement was observed in most of the cardinal and associated symptoms. Significant increase in peak expiratory flow rate, considerable decrease in absolute eosinophil count, and increased breath holding time were noticed. Withdrawal of modern emergency drugs, decreased frequency of attacks, improved quality of life were the major observations noticed in both groups. Conclusions: This study highlights the significance of traditional herbal formulations in noncommunicable diseases such as bronchial asthma, which can be used as an effective drug in place or along with modern drugs. PMID:27313413

  6. Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

    PubMed

    Pantchev, Nikola; Schnyder, Manuela; Vrhovec, Majda Globokar; Schaper, Roland; Tsachev, Ilia

    2015-08-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

  7. Comparison of coprological, immunological and molecular methods for the detection of dogs infected with Angiostrongylus vasorum before and after anthelmintic treatment.

    PubMed

    Schnyder, M; Jefferies, R; Schucan, A; Morgan, E R; Deplazes, P

    2015-09-01

    Timely diagnosis of the nematode Angiostrongylus vasorum in dogs is important in view of severe and permanent lung and cardiovascular lesions that may occur. The performance of the classical Baermann coprological method was compared with ELISAs for the serological detection of circulating antigen and specific antibodies and with Polymerase chain reaction (PCR) performed on EDTA blood, feces and tracheal swabs of serial samples from experimentally inoculated dogs over 13 weeks post inoculation (wpi) (n = 16) and following anthelmintic treatment (n = 6). Patency was observed from 6.7 to 7.6 wpi in all dogs, Baermann results were then mostly positive (116/119, 97%) during the patent period, with wide variations in the numbers of first stage larvae numbers. Blood PCR was tested positive on 1-2 occasions in 11/16 dogs in the pre-patent period, while all tested positive by antibody-detection ELISA by 6 wpi. The proportion of dogs testing positive by fecal PCR and antigen-detection ELISA rose early in the patent period. Tracheal swabs were occasionally DNA-positive in 3/16 dogs starting from 10 wpi. Following treatment, larval excretion stopped within 3 weeks and blood PCR results became negative within 1 week (5/6 dogs), while 4/6 dogs were positive for parasite DNA in tracheal swabs. Parasite antigen and specific antibodies both persisted in the blood for 3-9 weeks after treatment, with average optical densities and the proportion of positive dogs falling gradually, while results using other tests were much more variable. Results indicate that the earliest and most consistent results are obtained by the ELISAs, which can also be used for monitoring dogs after anthelmintic treatment.

  8. Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

    PubMed

    Pantchev, Nikola; Schnyder, Manuela; Vrhovec, Majda Globokar; Schaper, Roland; Tsachev, Ilia

    2015-08-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs. PMID:26152413

  9. Comparison of coprological, immunological and molecular methods for the detection of dogs infected with Angiostrongylus vasorum before and after anthelmintic treatment.

    PubMed

    Schnyder, M; Jefferies, R; Schucan, A; Morgan, E R; Deplazes, P

    2015-09-01

    Timely diagnosis of the nematode Angiostrongylus vasorum in dogs is important in view of severe and permanent lung and cardiovascular lesions that may occur. The performance of the classical Baermann coprological method was compared with ELISAs for the serological detection of circulating antigen and specific antibodies and with Polymerase chain reaction (PCR) performed on EDTA blood, feces and tracheal swabs of serial samples from experimentally inoculated dogs over 13 weeks post inoculation (wpi) (n = 16) and following anthelmintic treatment (n = 6). Patency was observed from 6.7 to 7.6 wpi in all dogs, Baermann results were then mostly positive (116/119, 97%) during the patent period, with wide variations in the numbers of first stage larvae numbers. Blood PCR was tested positive on 1-2 occasions in 11/16 dogs in the pre-patent period, while all tested positive by antibody-detection ELISA by 6 wpi. The proportion of dogs testing positive by fecal PCR and antigen-detection ELISA rose early in the patent period. Tracheal swabs were occasionally DNA-positive in 3/16 dogs starting from 10 wpi. Following treatment, larval excretion stopped within 3 weeks and blood PCR results became negative within 1 week (5/6 dogs), while 4/6 dogs were positive for parasite DNA in tracheal swabs. Parasite antigen and specific antibodies both persisted in the blood for 3-9 weeks after treatment, with average optical densities and the proportion of positive dogs falling gradually, while results using other tests were much more variable. Results indicate that the earliest and most consistent results are obtained by the ELISAs, which can also be used for monitoring dogs after anthelmintic treatment. PMID:26036271

  10. Efficacy and safety of imidacloprid/moxidectin spot-on solution and fenbendazole in the treatment of dogs naturally infected with Angiostrongylus vasorum (Baillet, 1866).

    PubMed

    Willesen, J L; Kristensen, A T; Jensen, A L; Heine, J; Koch, J

    2007-07-20

    A randomized, blinded, controlled multicentre field trial study was conducted to evaluate the efficacy and safety of imidacloprid 10%/moxidectin 2.5% spot-on solution and fenbendazole in treating dogs naturally infected with Angiostrongylus vasorum. Dogs were randomly treated either with a single dose of 0.1 ml/kg bodyweight of imidacloprid 10%/moxidectin 2.5% spot-on solution or with 25 mg/kg bodyweight fenbendazole per os for 20 days. The study period was 42 days with dogs being examined on days 0, 7 and 42. The primary efficacy parameter was the presence of L1 larvae in faecal samples evaluated by a Baermann test from three consecutive days. Thoracic radiographs performed on each visit were being taken as a paraclinical parameter to support the results of the Baermann test. Twenty-seven dogs in the imidacloprid/moxidectin group and 23 dogs in the fenbendazole group completed the study according to protocol. The efficacies of the two treatment protocols were 85.2% (imidacloprid/moxidectin) and 91.3% (fenbendazole) with no significant difference between treatment groups. On radiographic evaluation pulmonary parenchyma showed similar improvement in each group. No serious adverse effects to treatment were recorded: most of the minor adverse effects were gastrointestinal such as diarrhea (nine dogs), vomitus (eight dogs) and salivation (three dogs). In general, these adverse effects were of short duration (1-2 days) within the first few days after treatment start and required little or no treatment. This prospective study demonstrates that both treatment protocols used are efficacious under field conditions, that treatment of mildly to moderately infected dogs with either of these protocols is safe and yields an excellent prognosis for recovering from the infection.

  11. Zebrafish vasa homologue RNA is localized to the cleavage planes of 2- and 4-cell-stage embryos and is expressed in the primordial germ cells.

    PubMed

    Yoon, C; Kawakami, K; Hopkins, N

    1997-08-01

    Identification and manipulation of the germ line are important to the study of model organisms. Although zebrafish has recently emerged as a model for vertebrate development, the primordial germ cells (PGCs) in this organism have not been previously described. To identify a molecular marker for the zebrafish PGCs, we cloned the zebrafish homologue of the Drosophila vasa gene, which, in the fly, encodes a germ-cell-specific protein. Northern blotting revealed that zebrafish vasa homologue (vas) transcript is present in embryos just after fertilization, and hence it is probably maternally supplied. Using whole-mount in situ hybridization, we investigated the expression pattern of vas RNA in zebrafish embryos from the 1-cell stage to 10 days of development. Here we present evidence that vas RNA is a germ-cell-specific marker, allowing a description of the zebrafish PGCs for the first time. Furthermore, vas transcript was detected in a novel pattern, localized to the cleavage planes in 2- and 4-cell-stage embryos. During subsequent cleavages, the RNA is segregated as subcellular clumps to a small number of cells that may be the future germ cells. These results suggest new ways in which one might develop techniques for the genetic manipulation of zebrafish. Furthermore, they provide the basis for further studies on this novel RNA localization pattern and on germ-line development in general.

  12. Summary of cases of adventitial cystic disease of the popliteal artery.

    PubMed Central

    Flanigan, D P; Burnham, S J; Goodreau, J J; Bergan, J J

    1979-01-01

    Adventitial cystic disease of the popliteal artery is explored. The results of correspondence with authors reporting this condition are elaborated upon. This has provided an opportunity to discuss the history of the condition, the findings in 115 cases which have come to the attention of the Correspondence Office dealing with this entity, and the results of treatment. A discussion of the suspected etiology of the condition is presented. The condition remains one of unknown etiology which can be treated by cyst evacuation or aspiration when the popliteal artery is patent and which is best treated by arterial reconstruction when the artery is occluded. The results of such treatment are good but are dependent upon technical excellence of the operative procedure. PMID:426549

  13. Three-dimensional segmentation of luminal and adventitial borders in serial intravascular ultrasound images

    NASA Technical Reports Server (NTRS)

    Shekhar, R.; Cothren, R. M.; Vince, D. G.; Chandra, S.; Thomas, J. D.; Cornhill, J. F.

    1999-01-01

    Intravascular ultrasound (IVUS) provides exact anatomy of arteries, allowing accurate quantitative analysis. Automated segmentation of IVUS images is a prerequisite for routine quantitative analyses. We present a new three-dimensional (3D) segmentation technique, called active surface segmentation, which detects luminal and adventitial borders in IVUS pullback examinations of coronary arteries. The technique was validated against expert tracings by computing correlation coefficients (range 0.83-0.97) and William's index values (range 0.37-0.66). The technique was statistically accurate, robust to image artifacts, and capable of segmenting a large number of images rapidly. Active surface segmentation enabled geometrically accurate 3D reconstruction and visualization of coronary arteries and volumetric measurements.

  14. Acceleration of adventitious shoots by interaction between exogenous hormone and adenine sulphate in Althaea officinalis L.

    PubMed

    Naz, Ruphi; Anis, M

    2012-11-01

    In the current study attempts were made to investigate the effects of three different phases of callus induction followed by adventitious regeneration from leaf segments (central and lateral vein). Callus induction was observed in Murashige and Skoog's (MS) medium supplemented with 15.0 μM 2,4-dichloro phenoxy acetic acid (2,4-D). Adventitious shoot buds formation was achieved on MS medium supplemented with 7.5 μM 2,4-D and 20.0 μM AdS in liquid medium as it induced 19.2 ± 0.58 buds in central vein explants. Addition of different growth regulators (cytokinins-6-benzyladenine, kinetin and 2-isopentenyl adenine alone or in combination with auxins-indole-3-acetic acid, indole-3-butyric acid and α-naphthalene acetic acid, improved the shoot regeneration efficiency, in which 5.0 μM 6-benzyl adenine along with 0.25 μM α-naphthalene acetic acid was shown to be the most effective medium for maximum shoot regeneration (81.3 %) with 24.6 number of shoots and 4.4 ± 0.08 cm shoot length per explant. Leaf culture of central veins led to better shoot formation capacity in comparison to lateral vein. Rooting was readily achieved on the differentiated shoots on 1/2 MS medium augmented with 20.0 μM indole-3-butyric acid. The plants were successfully hardened off in sterile soilrite followed by their establishment in garden soil with 80 % survival rate.

  15. Adventitious Shoot Regeneration from Leaf Explant of Dwarf Hygro (Hygrophila polysperma (Roxb.) T. Anderson)

    PubMed Central

    Karataş, Mehmet; Aasim, Muhammad; Çınar, Ayşegül; Dogan, Muhammet

    2013-01-01

    Dwarf hygro (Hygrophila polysperma) is an ornamental aquatic plant that changes its leaf colours to pinkish in high light. It is listed as a medicinal plant in medicinal plant lists of Indian states of West Bengal and Karnataka. It is also used as a screening tool for toxicities and a bioindicator to detect and control algae. The study reported in vitro adventitious shoot regeneration from leaf explants cultured on MS medium containing 0.10–1.60 mg/L Kin/TDZ with or without 0.10 mg/L IBA and 500 mg/L Amoklavin to eradicate endogenic bacterial contamination. Direct adventitious shoot regeneration started within one week from both culture mediums followed by late callus induction which was more prominent on TDZ containing media compared to Kin containing media. Addition of 0.10 mg/L IBA with both Kin and TDZ increased shoot regeneration frequency, mean number of shoots per explant, and mean shoot length. Maximum number of 16.33 and 20.55 shoots per explant was obtained on MS medium containing 0.80 + 0.10 mg/L Kin-IBA and 0.10 + 0.10 mg/L TDZ-IBA, respectively. Regenerated shoots were rooted on MS medium containing 0.20–1.00 mg/L IBA followed by successfull acclimatization in aquariums. Regenerated plantlets were also tested in jars containing distilled water that showed the pH 6–9 for the best plant growth and development. PMID:23853539

  16. Adventitious shoot regeneration from leaf explant of dwarf hygro (Hygrophila polysperma (Roxb.) T. Anderson).

    PubMed

    Karataş, Mehmet; Aasim, Muhammad; Çınar, Ayşegül; Dogan, Muhammet

    2013-01-01

    Dwarf hygro (Hygrophila polysperma) is an ornamental aquatic plant that changes its leaf colours to pinkish in high light. It is listed as a medicinal plant in medicinal plant lists of Indian states of West Bengal and Karnataka. It is also used as a screening tool for toxicities and a bioindicator to detect and control algae. The study reported in vitro adventitious shoot regeneration from leaf explants cultured on MS medium containing 0.10-1.60 mg/L Kin/TDZ with or without 0.10 mg/L IBA and 500 mg/L Amoklavin to eradicate endogenic bacterial contamination. Direct adventitious shoot regeneration started within one week from both culture mediums followed by late callus induction which was more prominent on TDZ containing media compared to Kin containing media. Addition of 0.10 mg/L IBA with both Kin and TDZ increased shoot regeneration frequency, mean number of shoots per explant, and mean shoot length. Maximum number of 16.33 and 20.55 shoots per explant was obtained on MS medium containing 0.80 + 0.10 mg/L Kin-IBA and 0.10 + 0.10 mg/L TDZ-IBA, respectively. Regenerated shoots were rooted on MS medium containing 0.20-1.00 mg/L IBA followed by successfull acclimatization in aquariums. Regenerated plantlets were also tested in jars containing distilled water that showed the pH 6-9 for the best plant growth and development. PMID:23853539

  17. Estimating rat renal medullary interstitial oncotic pressures and the driving force for fluid uptake into ascending vasa recta

    PubMed Central

    MacPhee, P J

    1998-01-01

    We have investigated the potential contribution of medullary interstitial oncotic pressure to the net balance of forces influencing fluid movement through the walls of the ascending vasa recta (AVR) in the exposed papillae of 2-week-old Sprague-Dawley rats.Using a capillary zone electrophoresis (CZE) assay, hyaluronan (HA) concentrations were measured in fresh tissue slices from the renal papilla. HA content per wet weight of tissue decreased from tip to base of the papilla, but as a function of cell-free interstitial space (IS) HA concentration was relatively constant at 1.6 μg HA per mg IS up to 1600–1800 μm from the tip. Thereafter the concentration fell rapidly to near zero levels.The volume of the IS was determined using a transmission electron micrographic study of the papilla in age-matched rats. Total interstitial volume (i.e. IS + IC, the interstitial cell volume), as a function of total tissue volume, decreased only slightly between 0 and 1800 μm (i.e. from about 40 to 35%). IS and IC were found to be reciprocally related with IS decreasing from 21.8 to 10.2%, while IC increased from 18.3 to 25.2% over the 1800 μm.Total interstitial oncotic pressures were estimated as the sum of the oncotic pressure due to HA alone plus the oncotic pressure of albumin (A) in an HA matrix. Taking into account reflection coefficients to HA and A, there was an effective oncotic pressure (Eπ) of between 3.46 and 6.0 cmH2O on the interstitial side of the AVR. Under free flow conditions an Eπ in this range is sufficient to result in a net inward driving force of between 5.4 and 9.3 cmH2O, sufficient to account for current estimates of water reabsorption by the AVR.HA concentrations in the papilla increased over the first 3 weeks of life from 0.313 ±0.09 μg (mg wet weight of slice)−1 (mean ± s.e.m.) at 1 week to 0.563 ±0.06 at 3 weeks of age (P > 0.01), in parallel with an age-dependent increase in mean urine osmolarity. It is suggested that the increasing

  18. Foliar application of glyphosate affects molecular mechanisms in underground adventitious buds of leafy spurge (Euphorbia esula) and alters their vegetative growth patterns

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Long term control of leafy spurge with glyphosate requires multiple applications because the plant reproduces vegetatively from abundant underground adventitious buds (UABs). Determining the molecular mechanisms involved in controlling vegetative reproduction in leafy spurge following foliar glyphos...

  19. Adventitious roots of wheat seedlings that emerge in oxygen-deficient conditions have increased root diameters with highly developed lysigenous aerenchyma.

    PubMed

    Yamauchi, Takaki; Abe, Fumitaka; Kawaguchi, Kentaro; Oyanagi, Atsushi; Nakazono, Mikio

    2014-01-01

    Exposing roots of plants to hypoxic conditions is known to greatly improve their anoxic stress tolerance. We previously showed that pre-treatment of wheat seedlings with an ethylene precursor, 1-aminocyclopropanecarboxylic acid (ACC), enhanced their tolerance of oxygen-deficient conditions. Although ACC-pretreated seminal roots of wheat seedlings grown under oxygen-deficient conditions avoided root tip death, they elongated very little. In the present study, we assessed the effects of ethylene on the responses of adventitious roots of wheat seedlings to oxygen-deficient conditions. Lysigenous aerenchyma formation in the adventitious roots of wheat seedlings pretreated with ACC appeared to reduce tip death under oxygen-deficient conditions, but the adventitious roots, like the seminal roots, hardly elongated. We also found that adventitious roots that emerge in oxygen-deficient conditions continued to elongate even under such conditions. The adventitious roots emerged in oxygen-deficient conditions were found to have thicker root diameters than those emerged in aerated conditions. These results suggest that the adventitious roots with thicker root diameters can better cope with oxygen-deficient conditions. Measurements of the area of the lysigenous aerenchyma confirmed that the increased root diameters have a greater amount of air space generated by lysigenous aerenchyma formation.

  20. Adventitious roots of wheat seedlings that emerge in oxygen-deficient conditions have increased root diameters with highly developed lysigenous aerenchyma.

    PubMed

    Yamauchi, Takaki; Abe, Fumitaka; Kawaguchi, Kentaro; Oyanagi, Atsushi; Nakazono, Mikio

    2014-01-01

    Exposing roots of plants to hypoxic conditions is known to greatly improve their anoxic stress tolerance. We previously showed that pre-treatment of wheat seedlings with an ethylene precursor, 1-aminocyclopropanecarboxylic acid (ACC), enhanced their tolerance of oxygen-deficient conditions. Although ACC-pretreated seminal roots of wheat seedlings grown under oxygen-deficient conditions avoided root tip death, they elongated very little. In the present study, we assessed the effects of ethylene on the responses of adventitious roots of wheat seedlings to oxygen-deficient conditions. Lysigenous aerenchyma formation in the adventitious roots of wheat seedlings pretreated with ACC appeared to reduce tip death under oxygen-deficient conditions, but the adventitious roots, like the seminal roots, hardly elongated. We also found that adventitious roots that emerge in oxygen-deficient conditions continued to elongate even under such conditions. The adventitious roots emerged in oxygen-deficient conditions were found to have thicker root diameters than those emerged in aerated conditions. These results suggest that the adventitious roots with thicker root diameters can better cope with oxygen-deficient conditions. Measurements of the area of the lysigenous aerenchyma confirmed that the increased root diameters have a greater amount of air space generated by lysigenous aerenchyma formation. PMID:25764431

  1. Adventitious roots of wheat seedlings that emerge in oxygen-deficient conditions have increased root diameters with highly developed lysigenous aerenchyma

    PubMed Central

    Yamauchi, Takaki; Abe, Fumitaka; Kawaguchi, Kentaro; Oyanagi, Atsushi; Nakazono, Mikio

    2014-01-01

    Exposing roots of plants to hypoxic conditions is known to greatly improve their anoxic stress tolerance. We previously showed that pre-treatment of wheat seedlings with an ethylene precursor, 1-aminocyclopropanecarboxylic acid (ACC), enhanced their tolerance of oxygen-deficient conditions. Although ACC-pretreated seminal roots of wheat seedlings grown under oxygen-deficient conditions avoided root tip death, they elongated very little. In the present study, we assessed the effects of ethylene on the responses of adventitious roots of wheat seedlings to oxygen-deficient conditions. Lysigenous aerenchyma formation in the adventitious roots of wheat seedlings pretreated with ACC appeared to reduce tip death under oxygen-deficient conditions, but the adventitious roots, like the seminal roots, hardly elongated. We also found that adventitious roots that emerge in oxygen-deficient conditions continued to elongate even under such conditions. The adventitious roots emerged in oxygen-deficient conditions were found to have thicker root diameters than those emerged in aerated conditions. These results suggest that the adventitious roots with thicker root diameters can better cope with oxygen-deficient conditions. Measurements of the area of the lysigenous aerenchyma confirmed that the increased root diameters have a greater amount of air space generated by lysigenous aerenchyma formation. PMID:24690588

  2. Chemical characterization and prebiotic activity of fructo-oligosaccharides from Stevia rebaudiana (Bertoni) roots and in vitro adventitious root cultures.

    PubMed

    Sanches Lopes, Sheila Mara; Francisco, Mariane Grigio; Higashi, Bruna; de Almeida, Rafaela Takako Ribeiro; Krausová, Gabriela; Pilau, Eduardo Jorge; Gonçalves, José Eduardo; Gonçalves, Regina Aparecida Correia; Oliveira, Arildo José Braz de

    2016-11-01

    Stevia rebaudiana (Bertoni) is widely studied because of its foliar steviol glycosides. Fructan-type polysaccharides were recently isolated from its roots. Fructans are reserve carbohydrates that have important positive health effects and technological applications in the food industry. The objective of the present study was to isolate and characterize fructo-oligosaccharides (FOSs) from S. rebaudiana roots and in vitro adventitious root cultures and evaluate the potential prebiotic effect of these molecules. The in vitro adventitious root cultures were obtained using a roller bottle system. Chemical analyses (gas chromatography-mass spectrometry, (1)H nuclear magnetic resonance, and off-line electrospray ionization-mass spectrometry) revealed similar chemical properties of FOSs that were obtained from the different sources. The potential prebiotic effects of FOSs that were isolated from S. rebaudiana roots enhanced the growth of both bifidobacteria and lactobacilli, with strains specificity in their fermentation ability. PMID:27516323

  3. The effects of pruning and nodal adventitious roots on polychlorinated biphenyl uptake by Cucurbita pepo grown in field conditions.

    PubMed

    Low, Jennifer E; Åslund, Melissa L Whitfield; Rutter, Allison; Zeeb, Barbara A

    2011-03-01

    Two cultivation techniques (i-pruning and ii-nodal adventitious root encouragement) were investigated for their ability to increase PCB phytoextraction by Cucurbita pepo ssp pepo cv. Howden (pumpkin) plants in situ at a contaminated industrial site in Ontario (Aroclor 1248, mean soil [PCB] = 5.6 μg g(-1)). Pruning was implemented to increase plant biomass close to the root where PCB concentration is known to be highest. This treatment was found to have no effect on final shoot biomass or PCB concentration. However, material pruned from the plant is not included in the final shoot biomass. The encouragement of nodal adventitious roots at stem nodes did significantly increase the PCB concentration in the primary stem, while not affecting shoot biomass. Both techniques are easily applied cultivation practices that may be implemented to decrease phytoextraction treatment time.

  4. Pilot-scale culture of Hypericum perforatum L. adventitious roots in airlift bioreactors for the production of bioactive compounds.

    PubMed

    Cui, Xi-Hua; Murthy, Hosakatte Niranjana; Paek, Kee-Yoeup

    2014-09-01

    Hypericum perforatum L. (St. John's Wort) is an important medicinal plant which is widely used in the treatment for depression and irritable bowel syndrome. It is also used as a dietary supplement. Major bioactive phytochemicals of H. perforatum are phenolics and flavonoids. Quality of these phytochemicals is dramatically influenced by environmental and biological factors in the field grown plants. As an alternative, we have developed adventitious root cultures in large-scale bioreactors for the production of useful phytochemicals. Adventitious roots of H. perforatum were cultured in 500 l pilot-scale airlift bioreactors using half-strength Murashige and Skoog medium with an ammonium and nitrate ratio of 5:25 mM and supplemented with 1.0 mg l(-1) indole butyric acid, 0.1 mg l(-1) kinetin, and 3 % sucrose for the production of bioactive phenolics and flavonoids. Then 4.6 and 6.3 kg dry biomass were realized in the 500 l each of drum-type and balloon-type bioreactors, respectively. Accumulation of 66.9 mg g(-1) DW of total phenolics, 48.6 mg g(-1) DW of total flavonoids, 1.3 mg g(-1) DW of chlorogenic acid, 0.01 mg g(-1) DW of hyperin, 0.04 mg g(-1) DW of hypericin, and 0.01 mg g(-1) DW of quercetin could be achieved with adventitious roots cultured in 500 l balloon-type airlift bioreactors. Our findings demonstrate the possibilities of using H. perforatum adventitious root cultures for the production of useful phytochemicals to meet the demand of pharmaceutical and food industry.

  5. Pilot-scale culture of Hypericum perforatum L. adventitious roots in airlift bioreactors for the production of bioactive compounds.

    PubMed

    Cui, Xi-Hua; Murthy, Hosakatte Niranjana; Paek, Kee-Yoeup

    2014-09-01

    Hypericum perforatum L. (St. John's Wort) is an important medicinal plant which is widely used in the treatment for depression and irritable bowel syndrome. It is also used as a dietary supplement. Major bioactive phytochemicals of H. perforatum are phenolics and flavonoids. Quality of these phytochemicals is dramatically influenced by environmental and biological factors in the field grown plants. As an alternative, we have developed adventitious root cultures in large-scale bioreactors for the production of useful phytochemicals. Adventitious roots of H. perforatum were cultured in 500 l pilot-scale airlift bioreactors using half-strength Murashige and Skoog medium with an ammonium and nitrate ratio of 5:25 mM and supplemented with 1.0 mg l(-1) indole butyric acid, 0.1 mg l(-1) kinetin, and 3 % sucrose for the production of bioactive phenolics and flavonoids. Then 4.6 and 6.3 kg dry biomass were realized in the 500 l each of drum-type and balloon-type bioreactors, respectively. Accumulation of 66.9 mg g(-1) DW of total phenolics, 48.6 mg g(-1) DW of total flavonoids, 1.3 mg g(-1) DW of chlorogenic acid, 0.01 mg g(-1) DW of hyperin, 0.04 mg g(-1) DW of hypericin, and 0.01 mg g(-1) DW of quercetin could be achieved with adventitious roots cultured in 500 l balloon-type airlift bioreactors. Our findings demonstrate the possibilities of using H. perforatum adventitious root cultures for the production of useful phytochemicals to meet the demand of pharmaceutical and food industry. PMID:25096393

  6. Acetylcholinesterase-Inhibition and Antibacterial Activity of Mondia whitei Adventitious Roots and Ex vitro-Grown Somatic Embryogenic-Biomass

    PubMed Central

    Baskaran, Ponnusamy; Kumari, Aloka; Ncube, Bhekumthetho; Van Staden, Johannes

    2016-01-01

    Mondia whitei (Hook.f.) Skeels is an important endangered medicinal and commercial plant in South Africa. In vitro propagation systems are required for biomass production and bioactivity analysis to supplement wild resources/stocks. Adventitious roots from somatic embryogenic explants using suspension culture and ex vitro-grown plants produced via somatic embryogenesis were established using different plant growth regulator treatments. The adventitious root biomass and different parts of ex vitro-grown and mother plants were used to investigate the potential for acetylcholinesterase (AChE) and antibacterial activities. Adventitious roots derived from 2.5 μM indole-3-acetic acid (IAA) treatments and ex vitro-grown plants derived from meta-topolin riboside and IAA treatments gave the best AChE and antibacterial activities. The in vitro-established M. whitei and ex vitro biomass have comparable ability to function as inhibitors of acetylcholinesterase and antibacterial agents, and can be used as potent bioresources in traditional medicine. PMID:27752244

  7. Molecular cloning and expression analysis of the MTN gene during adventitious root development in IBA-induced tetraploid black locust.

    PubMed

    Quan, Jine; Zhang, Chunxia; Zhang, Sheng; Meng, Sen; Zhao, Zhong; Xu, Xuexuan

    2014-12-15

    5'-Methylthioadenosine (MTA) nucleosidase (MTN) plays a key role in the methionine (Met) recycling pathway of plants. Here, we report the isolation of the 1158 bp full-length, cDNA sequence encoding tetraploid black locust (Robinia pseudoacacia L.) MTN (TrbMTN), which contains an open reading frame of 810 bp that encodes a 269 amino acid protein. The amino acid sequence of TrbMTN has more than 88% sequence identity to the MTNs from other plants, with a closer phylogenetic relationship to MTNs from legumes than to MTNs from other plants. Subcellular localization analysis revealed that the TrbMTN gene localizes mainly to the cell membrane and cytoplasm of onion epidermal cells. Indole-3-butyric acid (IBA)-treated cuttings showed higher TrbMTN transcript levels than untreated control cuttings during root primordium and adventitious root formation. TrbMTN and key Met cycle genes showed differential expression in shoots, leaves, stems, and roots, with the highest expression observed in stems. IBA-treated cuttings also showed higher TrbMTN activity than control cuttings during root primordium and adventitious root formation. These results indicate that TrbMTN gene might play an important role in the regulation of IBA-induced adventitious root development in tetraploid black locust cuttings. PMID:25305345

  8. Identification of pectin methylesterase 3 as a basic pectin methylesterase isoform involved in adventitious rooting in Arabidopsis thaliana.

    PubMed

    Guénin, Stéphanie; Mareck, Alain; Rayon, Catherine; Lamour, Romain; Assoumou Ndong, Yves; Domon, Jean-Marc; Sénéchal, Fabien; Fournet, Françoise; Jamet, Elisabeth; Canut, Hervé; Percoco, Giuseppe; Mouille, Grégory; Rolland, Aurélia; Rustérucci, Christine; Guerineau, François; Van Wuytswinkel, Olivier; Gillet, Françoise; Driouich, Azeddine; Lerouge, Patrice; Gutierrez, Laurent; Pelloux, Jérôme

    2011-10-01

    • Here, we focused on the biochemical characterization of the Arabidopsis thaliana pectin methylesterase 3 gene (AtPME3; At3g14310) and its role in plant development. • A combination of biochemical, gene expression, Fourier transform-infrared (FT-IR) microspectroscopy and reverse genetics approaches were used. • We showed that AtPME3 is ubiquitously expressed in A. thaliana, particularly in vascular tissues. In cell wall-enriched fractions, only the mature part of the protein was identified, suggesting that it is processed before targeting the cell wall. In all the organs tested, PME activity was reduced in the atpme3-1 mutant compared with the wild type. This was related to the disappearance of an activity band corresponding to a pI of 9.6 revealed by a zymogram. Analysis of the cell wall composition showed that the degree of methylesterification (DM) of galacturonic acids was affected in the atpme3-1 mutant. A change in the number of adventitious roots was found in the mutant, which correlated with the expression of the gene in adventitious root primordia. • Our results enable the characterization of AtPME3 as a major basic PME isoform in A. thaliana and highlight its role in adventitious rooting.

  9. Integration of genetic, genomic and transcriptomic information identifies putative regulators of adventitious root formation in Populus

    DOE PAGES

    Ribeiro, Cintia L.; Silva, Cynthia M.; Drost, Derek R.; Novaes, Evandro; Novaes, Carolina R. D. B.; Dervinis, Christopher; Kirst, Matias

    2016-03-16

    In this study, adventitious roots (AR) develop from tissues other than the primary root, in a process physiologically regulated by phytohormones. Adventitious roots provide structural support and contribute to water and nutrient absorption, and are critical for commercial vegetative propagation of several crops. Here we quantified the number of AR, root architectural traits and root biomass in cuttings from a pseudo-backcross population of Populus deltoides and Populus trichocarpa. Quantitative trait loci (QTL) mapping and whole-transcriptome analysis of individuals with alternative QTL alleles for AR number were used to identify putative regulators of AR development. As a result, parental individuals andmore » progeny showed extensive segregation for AR developmental traits. Quantitative trait loci for number of AR mapped consistently in the same interval of linkage group (LG) II and LG XIV, explaining 7–10 % of the phenotypic variation. A time series transcriptome analysis identified 26,121 genes differentially expressed during AR development, particularly during the first 24 h after cuttings were harvested. Of those, 1929 genes were differentially regulated between individuals carrying alternative alleles for the two QTL for number of AR, in one or more time point. Eighty-one of these genes were physically located within the QTL intervals for number of AR, including putative homologs of the Arabidopsis genes SUPERROOT2 (SUR2) and TRYPTOPHAN SYNTHASE ALPHA CHAIN (TSA1), both of which are involved in the auxin indole-3-acetic acid (IAA) biosynthesis pathway. In conclusion, this study suggests the involvement of two genes of the tryptophan-dependent auxin biosynthesis pathway, SUR2 and TSA1, in the regulation of a critical trait for the clonal propagation of woody species. A possible model for this regulation is that poplar individuals that have poor AR formation synthesize auxin indole-3-acetic acid (IAA) primarily through the tryptophan (Trp) pathway. Much of

  10. The effects of melatonin on electrical field stimulation-evoked biphasic twitch responses in the ipsilateral and contralateral rat vasa deferentia after unilateral testicular torsion/detorsion.

    PubMed

    Barun, Süreyya; Ekingen, Gülşen; Mert Vural, Ismail; Türkyilmaz, Zafer; Başaklar, Can; Kale, Nuri; Sevim Ercan, Zeynep; Sarioğlu, Yusuf

    2005-05-01

    It is not known whether there is an impairment in vas deferens motility after unilateral testicular torsion/detorsion. Therefore, we aimed to determine whether the electrical field stimulation (EFS)-evoked biphasic contractions are altered in ipsilateral and contralateral rat vasa deferentia obtained from animals exposed to the unilateral testicular torsion/detorsion procedure. We also evaluated the effects of melatonin (MLT), which is a strong antioxidant, on these contractile responses. Rats were subjected to torsion of the left testis for 2 h and then detorsion was performed. Contractility studies were carried out 2 h or 24 h after detorsion. Vas deferens strips were prepared from both the ipsilateral and the contralateral site 2 h or 24 h after the detorsion procedure to record EFS-evoked biphasic twitch responses. The same experimental protocol was repeated for the MLT-treated rats. Both phases of EFS-evoked contractions were decreased after torsion/detorsion in the ipsilateral vas deferens. MLT treatment increased torsion/detorsion-induced reduction of both phases of contractions after 2 h and 24 h. In the contralateral vas deferens, the first phase of EFS-evoked contractions was not changed, while the second phase of contractions was diminished 2 h and 24 h after detorsion. Although MLT decreased the second phase of contractions 2 h and 24 h after detorsion, it reduced the first phase of contractions only 2 h after detorsion. These results suggest that MLT produces an inhibition on EFS-evoked biphasic twitch responses in the ipsilateral and contralateral rat vasa deferentia following unilateral testicular torsion/detorsion in the rat.

  11. The dietary significance of adventitious iron, zinc, copper and lead in domestically prepared food.

    PubMed

    Reilly, C

    1985-01-01

    The uptake of iron, zinc, copper and lead by food cooked under domestic conditions in utensils made of different metals (cast iron, aluminium, plain and tinned copper) was investigated. It was found that the metal content of the food was generally related to the metal in immediate contact with the food during cooking. Daily dietary intake could vary from 11 to 6 mg of iron, 11 to 9 mg of zinc, 2 to 1 mg of copper and 0.4 to 0.1 mg of lead, depending on the cooking utensils used. Dietary intake of the metals was also related to sources and domestic practices regarding water supply. Consistent use of municipal water from a domestic hot water system could contribute a daily intake of 32 mg iron, 29 mg zinc and 12 mg copper. Rainwater stored in a galvanized iron tank could provide 23 mg of zinc per day when used for domestic purposes. The nutritional and toxicological significance of such adventitious sources of metals in the diet are discussed. The need to consider them when investigating the metal intake of individuals is stressed.

  12. Excessive Adventitial Remodeling Leads to Early Aortic Maladaptation in Angiotensin-Induced Hypertension.

    PubMed

    Bersi, Matthew R; Bellini, Chiara; Wu, Jing; Montaniel, Kim R C; Harrison, David G; Humphrey, Jay D

    2016-05-01

    The primary function of central arteries is to store elastic energy during systole and to use it to sustain blood flow during diastole. Arterial stiffening compromises this normal mechanical function and adversely affects end organs, such as the brain, heart, and kidneys. Using an angiotensin II infusion model of hypertension in wild-type mice, we show that the thoracic aorta exhibits a dramatic loss of energy storage within 2 weeks that persists for at least 4 weeks. This diminished mechanical functionality results from increased structural stiffening as a result of an excessive accumulation of adventitial collagen, not a change in the intrinsic stiffness of the wall. A detailed analysis of the transmural biaxial wall stress suggests that the exuberant production of collagen results more from an inflammatory response than from a mechano-adaptation, hence reinforcing the need to control inflammation, not just blood pressure. Although most clinical assessments of arterial stiffening focus on intimal-medial thickening, these results suggest a need to measure and control the highly active and important adventitia.

  13. A Co-Opted Hormonal Cascade Activates Dormant Adventitious Root Primordia upon Flooding in Solanum dulcamara.

    PubMed

    Dawood, Thikra; Yang, Xinping; Visser, Eric J W; Te Beek, Tim A H; Kensche, Philip R; Cristescu, Simona M; Lee, Sangseok; Floková, Kristýna; Nguyen, Duy; Mariani, Celestina; Rieu, Ivo

    2016-04-01

    Soil flooding is a common stress factor affecting plants. To sustain root function in the hypoxic environment, flooding-tolerant plants may form new, aerenchymatous adventitious roots (ARs), originating from preformed, dormant primordia on the stem. We investigated the signaling pathway behind AR primordium reactivation in the dicot species Solanum dulcamara Transcriptome analysis indicated that flooding imposes a state of quiescence on the stem tissue, while increasing cellular activity in the AR primordia. Flooding led to ethylene accumulation in the lower stem region and subsequently to a drop in abscisic acid (ABA) level in both stem and AR primordia tissue. Whereas ABA treatment prevented activation of AR primordia by flooding, inhibition of ABA synthesis was sufficient to activate them in absence of flooding. Together, this reveals that there is a highly tissue-specific response to reduced ABA levels. The central role for ABA in the response differentiates the pathway identified here from the AR emergence pathway known from rice (Oryza sativa). Flooding and ethylene treatment also induced expression of the polar auxin transporter PIN2, and silencing of this gene or chemical inhibition of auxin transport inhibited primordium activation, even though ABA levels were reduced. Auxin treatment, however, was not sufficient for AR emergence, indicating that the auxin pathway acts in parallel with the requirement for ABA reduction. In conclusion, adaptation of S. dulcamara to wet habitats involved co-option of a hormonal signaling cascade well known to regulate shoot growth responses, to direct a root developmental program upon soil flooding. PMID:26850278

  14. [Impact of TDZ and NAA on adventitious bud induction and cluster bud multiplication in Tulipa edulis].

    PubMed

    Zhu, Li-Fang; Xu, Chao; Zhu, Zai-Biao; Yang, He-Tong; Guo, Qiao-Sheng; Xu, Hong-jian; Ma, Hong-Jian; Zhao, Gui-Hua

    2014-08-01

    To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established. PMID:25509282

  15. Percutaneous Image-Guided Aspiration and Sclerosis of Adventitial Cystic Disease of the Femoral Vein

    SciTech Connect

    Johnson, Jason M.; Kiankhooy, Armin; Bertges, Daniel J.; Morris, Christopher S.

    2009-07-15

    Adventitial cystic disease (ACD), also known as cystic mucoid or myxomatous degeneration, is a rare vascular disease mainly seen in arteries. Seventeen cases have been reported in the world literature. We report the first known case of ACD successfully treated with percutaneous image-guided ethanol sclerosis. Computed tomography showed a cystic mass adherent to the wall of the common femoral vein. An ultrasound examination revealed a deep venous thrombosis of the leg, secondary to extrinsic compression of the common femoral vein. Three years prior to our procedure, the cyst was aspirated, which partially relieved the patient's symptoms. Over the following 3 years the patient's symptoms worsened and a 10-cm discrepancy in thigh size developed, in addition to the deep venous thrombosis associated with lower-extremity edema. Using ultrasound guidance and fluoroscopic control, the cyst was drained and then sclerosed with absolute ethanol. The patient's symptoms and leg swelling resolved completely within several weeks. Follow-up physical examination and duplex ultrasound 6 months following sclerosis demonstrated resolution of the symptoms and elimination of the extrinsic compression effect of the ACD on the common femoral vein.

  16. Amide-conjugated indole-3-acetic acid and adventitious root formation in mung bean cuttings

    SciTech Connect

    Norcini, J.G.

    1986-01-01

    The purpose of this research was to investigate further the relationship between amide-conjugated auxin and adventitious root formation. Indoleacetylaspartic acid (IAA-aspartate) was positively identified as the predominant conjugate isolated from mung bean cuttings after the cuttings has been treated with 10/sup -3/ M IAA. In cuttings treated with (1-/sup 14/C)IAA immediately after excision (0 hr), the percent of extractable /sup 14/C in IAA-aspartate in the hypocotyl sharply increased until 36 hr, then steadily declined. (/sup 14/C)IAA was completely metabolized between 12 and 24 hr. The rooting activities of IAA-L-aspartate, IAA-L-alanine, and IAA-glycine were determined at various stages of root formation; some cuttings were pretreated with 10/sup -3/ M IAA at 0 hr. Pretreated cuttings that were treated with IAA-glycine at 12, 24, 36 hr exhibited the greatest consistency between replications, the greatest number of long roots, and the longest roots. The conjugates did not stimulate rooting as effectively as IAA, yet like IAA, generally enhanced rooting the greatest when applied before the first cell division (24 hr).

  17. Comparative transcriptional analysis provides new insights into the molecular basis of adventitious rooting recalcitrance in Eucalyptus.

    PubMed

    de Almeida, Márcia Rodrigues; de Bastiani, Daniela; Gaeta, Marcos Letaif; de Araújo Mariath, Jorge Ernesto; de Costa, Fernanda; Retallick, Jeffrey; Nolan, Lana; Tai, Helen H; Strömvik, Martina V; Fett-Neto, Arthur Germano

    2015-10-01

    Adventitious rooting (AR) is essential in clonal propagation. Eucalyptus globulus is relevant for the cellulose industry due to its low lignin content. However, several useful clones are recalcitrant to AR, often requiring exogenous auxin, adding cost to clonal garden operations. In contrast, E. grandis is an easy-to-root species widely used in clonal forestry. Aiming at contributing to the elucidation of recalcitrance causes in E. globulus, we conducted a comparative analysis with these two species differing in rooting competence, combining gene expression and anatomical techniques. Recalcitrance in E. globulus is reversed by exposure to exogenous indole-3-acetic acid (IAA), which promotes important gene expression modifications in both species. The endogenous content of IAA was significantly higher in E. grandis than in E. globulus. The cambium zone was identified as an active area during AR, concentrating the first cell divisions. Immunolocalization assay showed auxin accumulation in cambium cells, further indicating the importance of this region for rooting. We then performed a cambium zone-specific gene expression analysis during AR using laser microdissection. The results indicated that the auxin-related genes TOPLESS and IAA12/BODENLOS and the cytokinin-related gene ARR1may act as negative regulators of AR, possibly contributing to the hard-to-root phenotype of E. globulus. PMID:26398800

  18. Analysis of miRNAs and their targets during adventitious shoot organogenesis of Acacia crassicarpa.

    PubMed

    Liu, Weina; Yu, Wangning; Hou, Lingyu; Wang, Xiaoyu; Zheng, Fei; Wang, Weixuan; Liang, Di; Yang, Hailun; Jin, Yi; Xie, Xiangming

    2014-01-01

    Organogenesis is an important process for plant regeneration by tissue or cell mass differentiation to regenerate a complete plant. MicroRNAs (miRNAs) play an essential role in regulating plant development by mediating target genes at transcriptional and post-transcriptional levels, but the diversity of miRNAs and their potential roles in organogenesis of Acacia crassicarpa have rarely been investigated. In this study, approximately 10 million sequence reads were obtained from a small RNA library, from which 189 conserved miRNAs from 57 miRNA families, and 7 novel miRNAs from 5 families, were identified from A. crassicarpa organogenetic tissues. Target prediction for these miRNAs yielded 237 potentially unique genes, of which 207 received target Gene Ontology annotations. On the basis of a bioinformatic analysis, one novel and 13 conserved miRNAs were selected to investigate their possible roles in A. crassicarpa organogenesis by qRT-PCR. The stage-specific expression patterns of the miRNAs provided information on their possible regulatory functions, including shoot bud formation, modulated function after transfer of the culture to light, and regulatory roles during induction of organogenesis. This study is the first to investigate miRNAs associated with A. crassicarpa organogenesis. The results provide a foundation for further characterization of miRNA expression profiles and roles in the regulation of diverse physiological pathways during adventitious shoot organogenesis of A. crassicarpa.

  19. A Co-Opted Hormonal Cascade Activates Dormant Adventitious Root Primordia upon Flooding in Solanum dulcamara.

    PubMed

    Dawood, Thikra; Yang, Xinping; Visser, Eric J W; Te Beek, Tim A H; Kensche, Philip R; Cristescu, Simona M; Lee, Sangseok; Floková, Kristýna; Nguyen, Duy; Mariani, Celestina; Rieu, Ivo

    2016-04-01

    Soil flooding is a common stress factor affecting plants. To sustain root function in the hypoxic environment, flooding-tolerant plants may form new, aerenchymatous adventitious roots (ARs), originating from preformed, dormant primordia on the stem. We investigated the signaling pathway behind AR primordium reactivation in the dicot species Solanum dulcamara Transcriptome analysis indicated that flooding imposes a state of quiescence on the stem tissue, while increasing cellular activity in the AR primordia. Flooding led to ethylene accumulation in the lower stem region and subsequently to a drop in abscisic acid (ABA) level in both stem and AR primordia tissue. Whereas ABA treatment prevented activation of AR primordia by flooding, inhibition of ABA synthesis was sufficient to activate them in absence of flooding. Together, this reveals that there is a highly tissue-specific response to reduced ABA levels. The central role for ABA in the response differentiates the pathway identified here from the AR emergence pathway known from rice (Oryza sativa). Flooding and ethylene treatment also induced expression of the polar auxin transporter PIN2, and silencing of this gene or chemical inhibition of auxin transport inhibited primordium activation, even though ABA levels were reduced. Auxin treatment, however, was not sufficient for AR emergence, indicating that the auxin pathway acts in parallel with the requirement for ABA reduction. In conclusion, adaptation of S. dulcamara to wet habitats involved co-option of a hormonal signaling cascade well known to regulate shoot growth responses, to direct a root developmental program upon soil flooding.

  20. Rapid flooding-induced adventitious root development from preformed primordia in Solanum dulcamara.

    PubMed

    Dawood, Thikra; Rieu, Ivo; Wolters-Arts, Mieke; Derksen, Emiel B; Mariani, Celestina; Visser, Eric J W

    2014-01-01

    Flooding is a common stress factor in both natural and agricultural systems, and affects plant growth by the slow diffusion rate of gases in water. This results in low oxygen concentrations in submerged tissues, and hence in a decreased respiration rate. Understanding the responses of plants to flooding is essential for the management of wetland ecosystems, and may benefit research to improve the flood tolerance of crop species. This study describes the response to partial submergence of bittersweet (Solanum dulcamara). Bittersweet is a Eurasian species that grows both in dry habitats such as coastal dunes, and in wetlands, and therefore is a suitable model plant for studying responses to a variety of environmental stresses. A further advantage is that the species is closely related to flood-intolerant crops such as tomato and eggplant. The species constitutively develops dormant primordia on the stem, which we show to have a predetermined root identity. We investigated adventitious root growth from these primordia during flooding. The synchronized growth of roots from the primordia was detected after 2-3 days of flooding and was due to a combination of cell division and cell elongation. Gene expression analysis demonstrated that the molecular response to flooding began within 2 h and included activation of hypoxia and ethylene signalling genes. Unexpectedly, these early changes in gene expression were very similar in primordia and adjacent stem tissue, suggesting that there is a dominant general response in tissues during early flooding. PMID:24790121

  1. Excessive Adventitial Remodeling Leads to Early Aortic Maladaptation in Angiotensin-Induced Hypertension.

    PubMed

    Bersi, Matthew R; Bellini, Chiara; Wu, Jing; Montaniel, Kim R C; Harrison, David G; Humphrey, Jay D

    2016-05-01

    The primary function of central arteries is to store elastic energy during systole and to use it to sustain blood flow during diastole. Arterial stiffening compromises this normal mechanical function and adversely affects end organs, such as the brain, heart, and kidneys. Using an angiotensin II infusion model of hypertension in wild-type mice, we show that the thoracic aorta exhibits a dramatic loss of energy storage within 2 weeks that persists for at least 4 weeks. This diminished mechanical functionality results from increased structural stiffening as a result of an excessive accumulation of adventitial collagen, not a change in the intrinsic stiffness of the wall. A detailed analysis of the transmural biaxial wall stress suggests that the exuberant production of collagen results more from an inflammatory response than from a mechano-adaptation, hence reinforcing the need to control inflammation, not just blood pressure. Although most clinical assessments of arterial stiffening focus on intimal-medial thickening, these results suggest a need to measure and control the highly active and important adventitia. PMID:27001298

  2. [Impact of TDZ and NAA on adventitious bud induction and cluster bud multiplication in Tulipa edulis].

    PubMed

    Zhu, Li-Fang; Xu, Chao; Zhu, Zai-Biao; Yang, He-Tong; Guo, Qiao-Sheng; Xu, Hong-jian; Ma, Hong-Jian; Zhao, Gui-Hua

    2014-08-01

    To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established.

  3. Bioreactor with Ipomoea hederifolia adventitious roots and its endophyte Cladosporium cladosporioides for textile dye degradation.

    PubMed

    Patil, Swapnil M; Chandanshive, Vishal V; Rane, Niraj R; Khandare, Rahul V; Watharkar, Anuprita D; Govindwar, Sanjay P

    2016-04-01

    In vitro grown untransformed adventitious roots (AR) culture of Ipomoea hederifolia and its endophytic fungus (EF) Cladosporium cladosporioides decolorized Navy Blue HE2R (NB-HE2R) at a concentration of 20 ppm up to 83.3 and 65%, respectively within 96h. Whereas the AR-EF consortium decolorized the dye more efficiently and gave 97% removal within 36h. Significant inductions in the enzyme activities of lignin peroxidase, tyrosinase and laccase were observed in roots, while enzymes like tyrosinase, laccase and riboflavin reductase activities were induced in EF. Metabolites of dye were analyzed using UV-vis spectroscopy, FTIR and gas chromatography-mass spectrometry. Possible metabolic pathways of NB-HE2R were proposed with AR, EF and AR-EF systems independently. Looking at the superior efficacy of AR-EF system, a rhizoreactor was developed for the treatment of NB-HE2R at a concentration of 1000 ppm. Control reactor systems with independently grown AR and EF gave 94 and 85% NB-HE2R removal, respectively within 36h. The AR-EF rhizoreactor, however, gave 97% decolorization. The endophyte colonization additionally increased root and shoot lengths of candidate plants through mutualism. Combined bioreactor strategies can be effectively used for future eco-friendly remediation purposes. PMID:26803212

  4. Bioreactor with Ipomoea hederifolia adventitious roots and its endophyte Cladosporium cladosporioides for textile dye degradation.

    PubMed

    Patil, Swapnil M; Chandanshive, Vishal V; Rane, Niraj R; Khandare, Rahul V; Watharkar, Anuprita D; Govindwar, Sanjay P

    2016-04-01

    In vitro grown untransformed adventitious roots (AR) culture of Ipomoea hederifolia and its endophytic fungus (EF) Cladosporium cladosporioides decolorized Navy Blue HE2R (NB-HE2R) at a concentration of 20 ppm up to 83.3 and 65%, respectively within 96h. Whereas the AR-EF consortium decolorized the dye more efficiently and gave 97% removal within 36h. Significant inductions in the enzyme activities of lignin peroxidase, tyrosinase and laccase were observed in roots, while enzymes like tyrosinase, laccase and riboflavin reductase activities were induced in EF. Metabolites of dye were analyzed using UV-vis spectroscopy, FTIR and gas chromatography-mass spectrometry. Possible metabolic pathways of NB-HE2R were proposed with AR, EF and AR-EF systems independently. Looking at the superior efficacy of AR-EF system, a rhizoreactor was developed for the treatment of NB-HE2R at a concentration of 1000 ppm. Control reactor systems with independently grown AR and EF gave 94 and 85% NB-HE2R removal, respectively within 36h. The AR-EF rhizoreactor, however, gave 97% decolorization. The endophyte colonization additionally increased root and shoot lengths of candidate plants through mutualism. Combined bioreactor strategies can be effectively used for future eco-friendly remediation purposes.

  5. Influence of light and shoot development stage on leaf photosynthesis and carbohydrate status during the adventitious root formation in cuttings of Corylus avellana L.

    PubMed

    Tombesi, Sergio; Palliotti, Alberto; Poni, Stefano; Farinelli, Daniela

    2015-01-01

    Adventitious root formation in plant cuttings is influenced by many endogenous and environmental factors. Leaf photosynthesis during rooting of leafy cuttings in hard to root species can contribute to supply carbohydrates to the intensive metabolic processes related to adventious root formation. Light intensity during rooting is artificially kept low to decrease potential cutting desiccation, but can be limiting for photosynthetic activity. Furthermore, leafy cuttings collected from different part of the shoot can have a different ability to fuel adventitious root formation in cutting stem. The aim of this work was to determine the role of leaf photosynthesis on adventitious root formation in hazelnut (Corylus avellana L) (a hard-to-root specie) leafy cuttings and to investigate the possible influence of the shoot developmental stage on cutting rooting and survival in the post-rooting phase. Cutting rooting was closely related to carbohydrate content in cutting stems during the rooting process. Cutting carbohydrate status was positively influenced by leaf photosynthesis during rooting. Non-saturating light exposure of leafy cuttings can contribute to improve photosynthetic activity of leafy cuttings. Collection of cuttings from different part of the mother shoots influenced rooting percentage and this appear related to the different capability to concentrate soluble sugars in the cutting stem during rooting. Adventitious root formation depend on the carbohydrate accumulation at the base of the cutting. Mother shoot developmental stage and leaf photosynthesis appear pivotal factors for adventitious roots formation.

  6. Influence of light and shoot development stage on leaf photosynthesis and carbohydrate status during the adventitious root formation in cuttings of Corylus avellana L.

    PubMed Central

    Tombesi, Sergio; Palliotti, Alberto; Poni, Stefano; Farinelli, Daniela

    2015-01-01

    Adventitious root formation in plant cuttings is influenced by many endogenous and environmental factors. Leaf photosynthesis during rooting of leafy cuttings in hard to root species can contribute to supply carbohydrates to the intensive metabolic processes related to adventious root formation. Light intensity during rooting is artificially kept low to decrease potential cutting desiccation, but can be limiting for photosynthetic activity. Furthermore, leafy cuttings collected from different part of the shoot can have a different ability to fuel adventitious root formation in cutting stem. The aim of this work was to determine the role of leaf photosynthesis on adventitious root formation in hazelnut (Corylus avellana L) (a hard-to-root specie) leafy cuttings and to investigate the possible influence of the shoot developmental stage on cutting rooting and survival in the post-rooting phase. Cutting rooting was closely related to carbohydrate content in cutting stems during the rooting process. Cutting carbohydrate status was positively influenced by leaf photosynthesis during rooting. Non-saturating light exposure of leafy cuttings can contribute to improve photosynthetic activity of leafy cuttings. Collection of cuttings from different part of the mother shoots influenced rooting percentage and this appear related to the different capability to concentrate soluble sugars in the cutting stem during rooting. Adventitious root formation depend on the carbohydrate accumulation at the base of the cutting. Mother shoot developmental stage and leaf photosynthesis appear pivotal factors for adventitious roots formation. PMID:26635821

  7. An Efficient Method for Adventitious Root Induction from Stem Segments of Brassica Species

    PubMed Central

    Srikanth, Sandhya; Choong, Tsui Wei; Yan, An; He, Jie; Chen, Zhong

    2016-01-01

    Plant propagation via in vitro culture is a very laborious and time-consuming process. The growth cycle of some of the crop species is slow even in the field and the consistent commercial production is hard to maintain. Enhanced methods of reduced cost, materials and labor significantly impact the research and commercial production of field crops. In our studies, stem-segment explants of Brassica species were found to generate adventitious roots (AR) in aeroponic systems in less than a week. As such, the efficiency of rooting from stem explants of six cultivar varieties of Brassica spp was tested without using any plant hormones. New roots and shoots were developed from Brassica alboglabra (Kai Lan), B. oleracea var. acephala (purple kale), B. rapa L. ssp. chinensis L (Pai Tsai, Nai Bai C, and Nai Bai T) explants after 3 to 5 days of growing under 20 ± 2°C cool root zone temperature (C-RZT) and 4 to 7 days in 30 ± 2°C ambient root zone temperature (A-RZT). At the base of cut end, anticlinal and periclinal divisions of the cambial cells resulted in secondary xylem toward pith and secondary phloem toward cortex. The continuing mitotic activity of phloem parenchyma cells led to a ring of conspicuous white callus. Root initials formed from the callus which in turn developed into ARs. However, B. rapa var. nipposinica (Mizuna) explants were only able to root in C-RZT. All rooted explants were able to develop into whole plants, with higher biomass obtained from plants that grown in C-RZT. Moreover, explants from both RZTs produced higher biomass than plants grown from seeds (control plants). Rooting efficiency was affected by RZTs and explant cuttings of donor plants. Photosynthetic CO2 assimilation rate (Asat) and stomatal conductance (gssat) were significantly differentiated between plants derived from seeds and explants at both RZTs. All plants in A-RZT had highest transpiration rates. PMID:27446170

  8. When stress and development go hand in hand: main hormonal controls of adventitious rooting in cuttings

    PubMed Central

    da Costa, Cibele T.; de Almeida, Márcia R.; Ruedell, Carolina M.; Schwambach, Joseli; Maraschin, Felipe S.; Fett-Neto, Arthur G.

    2013-01-01

    Adventitious rooting (AR) is a multifactorial response leading to new roots at the base of stem cuttings, and the establishment of a complete and autonomous plant. AR has two main phases: (a) induction, with a requirement for higher auxin concentration; (b) formation, inhibited by high auxin and in which anatomical changes take place. The first stages of this process in severed organs necessarily include wounding and water stress responses which may trigger hormonal changes that contribute to reprogram target cells that are competent to respond to rooting stimuli. At severance, the roles of jasmonate and abscisic acid are critical for wound response and perhaps sink strength establishment, although their negative roles on the cell cycle may inhibit root induction. Strigolactones may also inhibit AR. A reduced concentration of cytokinins in cuttings results from the separation of the root system, whose tips are a relevant source of these root induction inhibitors. The combined increased accumulation of basipetally transported auxins from the shoot apex at the cutting base is often sufficient for AR in easy-to-root species. The role of peroxidases and phenolic compounds in auxin catabolism may be critical at these early stages right after wounding. The events leading to AR strongly depend on mother plant nutritional status, both in terms of minerals and carbohydrates, as well as on sink establishment at cutting bases. Auxins play a central role in AR. Auxin transporters control auxin canalization to target cells. There, auxins act primarily through selective proteolysis and cell wall loosening, via their receptor proteins TIR1 (transport inhibitor response 1) and ABP1 (Auxin-Binding Protein 1). A complex microRNA circuitry is involved in the control of auxin response factors essential for gene expression in AR. After root establishment, new hormonal controls take place, with auxins being required at lower concentrations for root meristem maintenance and cytokinins

  9. An Efficient Method for Adventitious Root Induction from Stem Segments of Brassica Species.

    PubMed

    Srikanth, Sandhya; Choong, Tsui Wei; Yan, An; He, Jie; Chen, Zhong

    2016-01-01

    Plant propagation via in vitro culture is a very laborious and time-consuming process. The growth cycle of some of the crop species is slow even in the field and the consistent commercial production is hard to maintain. Enhanced methods of reduced cost, materials and labor significantly impact the research and commercial production of field crops. In our studies, stem-segment explants of Brassica species were found to generate adventitious roots (AR) in aeroponic systems in less than a week. As such, the efficiency of rooting from stem explants of six cultivar varieties of Brassica spp was tested without using any plant hormones. New roots and shoots were developed from Brassica alboglabra (Kai Lan), B. oleracea var. acephala (purple kale), B. rapa L. ssp. chinensis L (Pai Tsai, Nai Bai C, and Nai Bai T) explants after 3 to 5 days of growing under 20 ± 2°C cool root zone temperature (C-RZT) and 4 to 7 days in 30 ± 2°C ambient root zone temperature (A-RZT). At the base of cut end, anticlinal and periclinal divisions of the cambial cells resulted in secondary xylem toward pith and secondary phloem toward cortex. The continuing mitotic activity of phloem parenchyma cells led to a ring of conspicuous white callus. Root initials formed from the callus which in turn developed into ARs. However, B. rapa var. nipposinica (Mizuna) explants were only able to root in C-RZT. All rooted explants were able to develop into whole plants, with higher biomass obtained from plants that grown in C-RZT. Moreover, explants from both RZTs produced higher biomass than plants grown from seeds (control plants). Rooting efficiency was affected by RZTs and explant cuttings of donor plants. Photosynthetic CO2 assimilation rate (Asat ) and stomatal conductance (gssat ) were significantly differentiated between plants derived from seeds and explants at both RZTs. All plants in A-RZT had highest transpiration rates.

  10. Auxin and cytokinin control formation of the quiescent centre in the adventitious root apex of arabidopsis

    PubMed Central

    Della Rovere, F.; Fattorini, L.; D'Angeli, S.; Veloccia, A.; Falasca, G.; Altamura, M. M.

    2013-01-01

    Background and Aims Adventitious roots (ARs) are part of the root system in numerous plants, and are required for successful micropropagation. In the Arabidopsis thaliana primary root (PR) and lateral roots (LRs), the quiescent centre (QC) in the stem cell niche of the meristem controls apical growth with the involvement of auxin and cytokinin. In arabidopsis, ARs emerge in planta from the hypocotyl pericycle, and from different tissues in in vitro cultured explants, e.g. from the stem endodermis in thin cell layer (TCL) explants. The aim of this study was to investigate the establishment and maintenance of the QC in arabidopsis ARs, in planta and in TCL explants, because information about this process is still lacking, and it has potential use for biotechnological applications. Methods Expression of PR/LR QC markers and auxin influx (LAX3)/efflux (PIN1) genes was investigated in the presence/absence of exogenous auxin and cytokinin. Auxin was monitored by the DR5::GUS system and cytokinin by immunolocalization. The expression of the auxin-biosynthetic YUCCA6 gene was also investigated by in situ hybridization in planta and in AR-forming TCLs from the indole acetic acid (IAA)-overproducing superroot2-1 mutant and its wild type. Key Results The accumulation of auxin and the expression of the QC marker WOX5 characterized the early derivatives of the AR founder cells, in planta and in in vitro cultured TCLs. By determination of PIN1 auxin efflux carrier and LAX3 auxin influx carrier activities, an auxin maximum was determined to occur at the AR tip, to which WOX5 expression was restricted, establishing the positioning of the QC. Cytokinin caused a restriction of LAX3 and PIN1 expression domains, and concomitantly the auxin biosynthesis YUCCA6 gene was expressed in the apex. Conclusions In ARs formed in planta and TCLs, the QC is established in a similar way, and auxin transport and biosynthesis are involved through cytokinin tuning. PMID:24061489

  11. An Efficient Method for Adventitious Root Induction from Stem Segments of Brassica Species.

    PubMed

    Srikanth, Sandhya; Choong, Tsui Wei; Yan, An; He, Jie; Chen, Zhong

    2016-01-01

    Plant propagation via in vitro culture is a very laborious and time-consuming process. The growth cycle of some of the crop species is slow even in the field and the consistent commercial production is hard to maintain. Enhanced methods of reduced cost, materials and labor significantly impact the research and commercial production of field crops. In our studies, stem-segment explants of Brassica species were found to generate adventitious roots (AR) in aeroponic systems in less than a week. As such, the efficiency of rooting from stem explants of six cultivar varieties of Brassica spp was tested without using any plant hormones. New roots and shoots were developed from Brassica alboglabra (Kai Lan), B. oleracea var. acephala (purple kale), B. rapa L. ssp. chinensis L (Pai Tsai, Nai Bai C, and Nai Bai T) explants after 3 to 5 days of growing under 20 ± 2°C cool root zone temperature (C-RZT) and 4 to 7 days in 30 ± 2°C ambient root zone temperature (A-RZT). At the base of cut end, anticlinal and periclinal divisions of the cambial cells resulted in secondary xylem toward pith and secondary phloem toward cortex. The continuing mitotic activity of phloem parenchyma cells led to a ring of conspicuous white callus. Root initials formed from the callus which in turn developed into ARs. However, B. rapa var. nipposinica (Mizuna) explants were only able to root in C-RZT. All rooted explants were able to develop into whole plants, with higher biomass obtained from plants that grown in C-RZT. Moreover, explants from both RZTs produced higher biomass than plants grown from seeds (control plants). Rooting efficiency was affected by RZTs and explant cuttings of donor plants. Photosynthetic CO2 assimilation rate (Asat ) and stomatal conductance (gssat ) were significantly differentiated between plants derived from seeds and explants at both RZTs. All plants in A-RZT had highest transpiration rates. PMID:27446170

  12. Progression Rates of Carotid Intima-media Thickness and Adventitial Diameter during the Menopausal Transition

    PubMed Central

    El Khoudary, Samar R.; Wildman, Rachel P.; Matthews, Karen; Thurston, Rebecca C.; Bromberger, Joyce T.; Sutton-Tyrrell, Kim

    2012-01-01

    Objectives The authors assessed whether the levels and progression rates of carotid intima-media thickness (IMT) and adventitial diameter (AD) vary by menopausal stage. Methods 249 Women (42–57 years old, premenopausal (49%) or early peri-menopausal (46%)) from the Study of Women’s Health Across the Nation were included in the current analysis. Participants were followed for up to 9 years (median=3.7 years) and had up to 5 carotid scans. Linear mixed models were used for analysis. Results The overall rate of change in IMT was 0.007 mm/year. Independent of age and race, progression rate of IMT increased substantially in late peri-menopausal stage (0.017 mm/year) compared to both premenopausal (0.007 mm/year) and early peri-menopausal (0.005 mm/year) stages; (P≤0.05). For AD, while the overall rate of change was negative (−0.009 mm/year), significant positive increases in the rate of change were observed in late peri-menopausal (0.024 mm/year) and postmenopausal (0.018 mm/year) stages compared to premenopausal stage (−0.032 mm/year); (P<0.05). In final models, postmenopausal stage was independently associated with higher levels of IMT and AD (P<0.05) compared to premenopausal stage. Conclusions During the menopausal transition, the carotid artery undergoes an adaptation that is reflected in adverse changes in IMT and AD. These changes may impact the vulnerability of the vessel to disease in older women. PMID:22990755

  13. Partial flooding enhances aeration in adventitious roots of black willow (Salix nigra) cuttings.

    PubMed

    Li, Shuwen; Reza Pezeshki, S; Douglas Shields, F

    2006-04-01

    Black willow (Salix nigra) cuttings are used for streambank stabilization where they are subjected to a range of soil moisture conditions including flooding. Flooding has been shown to adversely impact cutting performance, and improved understanding of natural adaptations to flooding might suggest handling and planting techniques to enhance success. However, data assessing the root aeration in adventitious roots that are developed on cuttings of woody species are scant. In addition, it appears that no data are available regarding aeration of the root system under partially flooded conditions. This experiment was designed to examine the effects of continuous flooding (CF) and partial flooding (PF) on aerenchyma formation and radial oxygen loss (ROL) in black willow cuttings. Photosynthetic and growth responses to these conditions were also investigated. Under laboratory condition, replicated potted cuttings were subjected to three treatments: no flooding (control, C), CF, and PF. Water was maintained above the soil surface in CF and at 10 cm depth in PF. Results indicated that after the 28-d treatments, root porosity ranged between 28.6% and 33.0% for the CF and C plants but was greater for the PF plants (39.2% for the drained and 37.2% for the flooded portions). A similar response pattern was found for ROL. In addition, CF treatment led to decreases in final root biomass and root/shoot ratio. Neither CF nor PF had any detectable adverse effects on plant gas exchange or photosystem II functioning. Our results indicated that S. nigra cuttings exhibited avoidance mechanisms in response to flooding, especially the partially flooded condition which is the most common occurrence in riparian systems.

  14. Quantitative risk assessment relating to adventitious presence of allergens in food: a probabilistic model applied to peanut in chocolate.

    PubMed

    Rimbaud, Loup; Heraud, Fanny; La Vieille, Sébastien; Leblanc, Jean-Charles; Crepet, Amélie

    2010-01-01

    Peanut allergy is a public health concern, owing to the high prevalence in France and the severity of the reactions. Despite peanut-containing product avoidance diets, a risk may exist due to the adventitious presence of peanut allergens in a wide range of food products. Peanut is not mentioned in their ingredients list, but precautionary labeling is often present. A method of quantifying the risk of allergic reactions following the consumption of such products is developed, taking the example of peanut in chocolate tablets. The occurrence of adventitious peanut proteins in chocolate and the dose-response relationship are estimated with a Bayesian approach using available published data. The consumption pattern is described by the French individual consumption survey INCA2. Risk simulations are performed using second-order Monte Carlo simulations, which separately propagates variability and uncertainty of the model input variables. Peanut allergens occur in approximately 36% of the chocolates, leading to a mean exposure level of 0.2 mg of peanut proteins per eating occasion. The estimated risk of reaction averages 0.57% per eating occasion for peanut-allergic adults. The 95% values of the risk stand between 0 and 3.61%, which illustrates the risk variability. The uncertainty, represented by the 95% credible intervals, is concentrated around these risk estimates. Children have similar results. The conclusion is that adventitious peanut allergens induce a risk of reaction for a part of the French peanut-allergic population. The method developed can be generalized to assess the risk due to the consumption of every foodstuff potentially contaminated by allergens.

  15. High Spatial Resolution MRI of Cystic Adventitial Disease of the Iliofemoral Vein Communicating with the Hip Joint

    SciTech Connect

    Michaelides, Michael; Pantziara, Maria Ioannidis, Kleanthis

    2013-05-14

    Venous cystic adventitial disease (CAD) is an extremely rare entity, and so far less than 20 cases have been described in the literature. Herein, we describe the imaging findings of CAD of iliofemoral vein in a 51-year-old woman who presented with leg swelling with special emphasis on high spatial resolution MRI, which demonstrated communication of the cyst with the hip joint. To our knowledge, this is the first description of high spatial resolution MRI findings in venous CAD supporting a new theory about the pathogenesis of venous CAD.

  16. Efficient uptake of blood-borne BK and JC polyomavirus-like particles in endothelial cells of liver sinusoids and renal vasa recta.

    PubMed

    Simon-Santamaria, Jaione; Rinaldo, Christine Hanssen; Kardas, Piotr; Li, Ruomei; Malovic, Ivana; Elvevold, Kjetil; McCourt, Peter; Smedsrød, Bård; Hirsch, Hans H; Sørensen, Karen Kristine

    2014-01-01

    Liver sinusoidal endothelial cells (LSECs) are specialized scavenger cells that mediate high-capacity clearance of soluble waste macromolecules and colloid material, including blood-borne adenovirus. To explore if LSECs function as a sink for other viruses in blood, we studied the fate of virus-like particles (VLPs) of two ubiquitous human DNA viruses, BK and JC polyomavirus, in mice. Like complete virions, VLPs specifically bind to receptors and enter cells, but unlike complete virions, they cannot replicate. 125I-labeled VLPs were used to assess blood decay, organ-, and hepatocellular distribution of ligand, and non-labeled VLPs to examine cellular uptake by immunohisto- and -cytochemistry. BK- and JC-VLPs rapidly distributed to liver, with lesser uptake in kidney and spleen. Liver uptake was predominantly in LSECs. Blood half-life (∼1 min), and tissue distribution of JC-VLPs and two JC-VLP-mutants (L55F and S269F) that lack sialic acid binding affinity, were similar, indicating involvement of non-sialic acid receptors in cellular uptake. Liver uptake was not mediated by scavenger receptors. In spleen, the VLPs localized to the red pulp marginal zone reticuloendothelium, and in kidney to the endothelial lining of vasa recta segments, and the transitional epithelium of renal pelvis. Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta. The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia. We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation. In addition, we report the novel finding that a subpopulation of endothelial cells in kidney, the main organ of polyomavirus persistence, showed selective and

  17. Efficient Uptake of Blood-Borne BK and JC Polyomavirus-Like Particles in Endothelial Cells of Liver Sinusoids and Renal Vasa Recta

    PubMed Central

    Simon-Santamaria, Jaione; Rinaldo, Christine Hanssen; Kardas, Piotr; Li, Ruomei; Malovic, Ivana; Elvevold, Kjetil; McCourt, Peter; Smedsrød, Bård

    2014-01-01

    Liver sinusoidal endothelial cells (LSECs) are specialized scavenger cells that mediate high-capacity clearance of soluble waste macromolecules and colloid material, including blood-borne adenovirus. To explore if LSECs function as a sink for other viruses in blood, we studied the fate of virus-like particles (VLPs) of two ubiquitous human DNA viruses, BK and JC polyomavirus, in mice. Like complete virions, VLPs specifically bind to receptors and enter cells, but unlike complete virions, they cannot replicate. 125I-labeled VLPs were used to assess blood decay, organ-, and hepatocellular distribution of ligand, and non-labeled VLPs to examine cellular uptake by immunohisto- and -cytochemistry. BK- and JC-VLPs rapidly distributed to liver, with lesser uptake in kidney and spleen. Liver uptake was predominantly in LSECs. Blood half-life (∼1 min), and tissue distribution of JC-VLPs and two JC-VLP-mutants (L55F and S269F) that lack sialic acid binding affinity, were similar, indicating involvement of non-sialic acid receptors in cellular uptake. Liver uptake was not mediated by scavenger receptors. In spleen, the VLPs localized to the red pulp marginal zone reticuloendothelium, and in kidney to the endothelial lining of vasa recta segments, and the transitional epithelium of renal pelvis. Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta. The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia. We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation. In addition, we report the novel finding that a subpopulation of endothelial cells in kidney, the main organ of polyomavirus persistence, showed selective and

  18. Nitric oxide and hydrogen peroxide alleviate drought stress in marigold explants and promote its adventitious root development.

    PubMed

    Liao, Wei-Biao; Huang, Gao-Bao; Yu, Ji-Hua; Zhang, Mei-Ling

    2012-09-01

    Drought stress is one of the most important environmental factors that regulates plant growth and development. In this study, we examined the effects of nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) on adventitious rooting in marigold (Tagetes erecta L.) under drought stress. The results showed that the promoting effect of NO or H(2)O(2) on rooting under drought stress was dose-dependent, with a maximal biological response at 10 μM NO donor sodium nitroprusside (SNP) or 600 μM H(2)O(2). Results also indicated that endogenous NO and H(2)O(2) may play crucial roles in rooting under drought conditions, and H(2)O(2) may be involved in rooting promoted by NO under drought stress. NO or H(2)O(2) treatment attenuated the destruction of mesophyll cells ultrastructure by drought stress. Similarly, NO or H(2)O(2) increased leaf chlorophyll content, chlorophyll fluorescence parameters (Fv/Fm, ΦPS II and qP), and hypocotyls soluble carbohydrate and protein content, while decreasing starch content. Results suggest that the protection of mesophyll cells ultrastructure by NO or H(2)O(2) under drought conditions improves the photosynthetic performance of leaves and alleviates the negative effects of drought on carbohydrate and nitrogen accumulation in explants, thereby adventitious rooting being promoted. PMID:22771430

  19. Plant Hormone Homeostasis, Signaling, and Function during Adventitious Root Formation in Cuttings

    PubMed Central

    Druege, Uwe; Franken, Philipp; Hajirezaei, Mohammad R.

    2016-01-01

    Adventitious root (AR) formation in cuttings is a multiphase developmental process, resulting from wounding at the cutting site and isolation from the resource and signal network of the whole plant. Though, promotive effects of auxins are widely used for clonal plant propagation, the regulation and function of plant hormones and their intricate signaling networks during AR formation in cuttings are poorly understood. In this focused review, we discuss our recent publications on the involvement of polar auxin transport (PAT) and transcriptional regulation of auxin and ethylene action during AR formation in petunia cuttings in a broad context. Integrating new findings on cuttings of other plant species and general models on plant hormone networks, a model on the regulation and function of auxin, ethylene, and jasmonate in AR formation of cuttings is presented. PAT and cutting off from the basipetal auxin drain are considered as initial principles generating early accumulation of IAA in the rooting zone. This is expected to trigger a self-regulatory process of auxin canalization and maximization to responding target cells, there inducing the program of AR formation. Regulation of auxin homeostasis via auxin influx and efflux carriers, GH3 proteins and peroxidases, of flavonoid metabolism, and of auxin signaling via AUX/IAA proteins, TOPLESS, ARFs, and SAUR-like proteins are postulated as key processes determining the different phases of AR formation. NO and H2O2 mediate auxin signaling via the cGMP and MAPK cascades. Transcription factors of the GRAS-, AP2/ERF-, and WOX-families link auxin signaling to cell fate specification. Cyclin-mediated governing of the cell cycle, modifications of sugar metabolism and microtubule and cell wall remodeling are considered as important implementation processes of auxin function. Induced by the initial wounding and other abiotic stress factors, up-regulation of ethylene biosynthesis, and signaling via ERFs and early accumulation of

  20. Plant Hormone Homeostasis, Signaling, and Function during Adventitious Root Formation in Cuttings.

    PubMed

    Druege, Uwe; Franken, Philipp; Hajirezaei, Mohammad R

    2016-01-01

    Adventitious root (AR) formation in cuttings is a multiphase developmental process, resulting from wounding at the cutting site and isolation from the resource and signal network of the whole plant. Though, promotive effects of auxins are widely used for clonal plant propagation, the regulation and function of plant hormones and their intricate signaling networks during AR formation in cuttings are poorly understood. In this focused review, we discuss our recent publications on the involvement of polar auxin transport (PAT) and transcriptional regulation of auxin and ethylene action during AR formation in petunia cuttings in a broad context. Integrating new findings on cuttings of other plant species and general models on plant hormone networks, a model on the regulation and function of auxin, ethylene, and jasmonate in AR formation of cuttings is presented. PAT and cutting off from the basipetal auxin drain are considered as initial principles generating early accumulation of IAA in the rooting zone. This is expected to trigger a self-regulatory process of auxin canalization and maximization to responding target cells, there inducing the program of AR formation. Regulation of auxin homeostasis via auxin influx and efflux carriers, GH3 proteins and peroxidases, of flavonoid metabolism, and of auxin signaling via AUX/IAA proteins, TOPLESS, ARFs, and SAUR-like proteins are postulated as key processes determining the different phases of AR formation. NO and H2O2 mediate auxin signaling via the cGMP and MAPK cascades. Transcription factors of the GRAS-, AP2/ERF-, and WOX-families link auxin signaling to cell fate specification. Cyclin-mediated governing of the cell cycle, modifications of sugar metabolism and microtubule and cell wall remodeling are considered as important implementation processes of auxin function. Induced by the initial wounding and other abiotic stress factors, up-regulation of ethylene biosynthesis, and signaling via ERFs and early accumulation of

  1. De Novo Characterization of the Mung Bean Transcriptome and Transcriptomic Analysis of Adventitious Rooting in Seedlings Using RNA-Seq

    PubMed Central

    Li, Shi-Weng; Shi, Rui-Fang; Leng, Yan

    2015-01-01

    Adventitious rooting is the most important mechanism underlying vegetative propagation and an important strategy for plant propagation under environmental stress. The present study was conducted to obtain transcriptomic data and examine gene expression using RNA-Seq and bioinformatics analysis, thereby providing a foundation for understanding the molecular mechanisms controlling adventitious rooting. Three cDNA libraries constructed from mRNA samples from mung bean hypocotyls during adventitious rooting were sequenced. These three samples generated a total of 73 million, 60 million, and 59 million 100-bp reads, respectively. These reads were assembled into 78,697 unigenes with an average length of 832 bp, totaling 65 Mb. The unigenes were aligned against six public protein databases, and 29,029 unigenes (36.77%) were annotated using BLASTx. Among them, 28,225 (35.75%) and 28,119 (35.62%) unigenes had homologs in the TrEMBL and NCBI non-redundant (Nr) databases, respectively. Of these unigenes, 21,140 were assigned to gene ontology classes, and a total of 11,990 unigenes were classified into 25 KOG functional categories. A total of 7,357 unigenes were annotated to 4,524 KOs, and 4,651 unigenes were mapped onto 342 KEGG pathways using BLAST comparison against the KEGG database. A total of 11,717 unigenes were differentially expressed (fold change>2) during the root induction stage, with 8,772 unigenes down-regulated and 2,945 unigenes up-regulated. A total of 12,737 unigenes were differentially expressed during the root initiation stage, with 9,303 unigenes down-regulated and 3,434 unigenes up-regulated. A total of 5,334 unigenes were differentially expressed between the root induction and initiation stage, with 2,167 unigenes down-regulated and 3,167 unigenes up-regulated. qRT-PCR validation of the 39 genes with known functions indicated a strong correlation (92.3%) with the RNA-Seq data. The GO enrichment, pathway mapping, and gene expression profiles reveal

  2. De Novo Characterization of the Mung Bean Transcriptome and Transcriptomic Analysis of Adventitious Rooting in Seedlings Using RNA-Seq.

    PubMed

    Li, Shi-Weng; Shi, Rui-Fang; Leng, Yan

    2015-01-01

    Adventitious rooting is the most important mechanism underlying vegetative propagation and an important strategy for plant propagation under environmental stress. The present study was conducted to obtain transcriptomic data and examine gene expression using RNA-Seq and bioinformatics analysis, thereby providing a foundation for understanding the molecular mechanisms controlling adventitious rooting. Three cDNA libraries constructed from mRNA samples from mung bean hypocotyls during adventitious rooting were sequenced. These three samples generated a total of 73 million, 60 million, and 59 million 100-bp reads, respectively. These reads were assembled into 78,697 unigenes with an average length of 832 bp, totaling 65 Mb. The unigenes were aligned against six public protein databases, and 29,029 unigenes (36.77%) were annotated using BLASTx. Among them, 28,225 (35.75%) and 28,119 (35.62%) unigenes had homologs in the TrEMBL and NCBI non-redundant (Nr) databases, respectively. Of these unigenes, 21,140 were assigned to gene ontology classes, and a total of 11,990 unigenes were classified into 25 KOG functional categories. A total of 7,357 unigenes were annotated to 4,524 KOs, and 4,651 unigenes were mapped onto 342 KEGG pathways using BLAST comparison against the KEGG database. A total of 11,717 unigenes were differentially expressed (fold change>2) during the root induction stage, with 8,772 unigenes down-regulated and 2,945 unigenes up-regulated. A total of 12,737 unigenes were differentially expressed during the root initiation stage, with 9,303 unigenes down-regulated and 3,434 unigenes up-regulated. A total of 5,334 unigenes were differentially expressed between the root induction and initiation stage, with 2,167 unigenes down-regulated and 3,167 unigenes up-regulated. qRT-PCR validation of the 39 genes with known functions indicated a strong correlation (92.3%) with the RNA-Seq data. The GO enrichment, pathway mapping, and gene expression profiles reveal

  3. Accounting for nanometer-thick adventitious carbon contamination in X-ray absorption spectra of carbon-based materials.

    PubMed

    Mangolini, Filippo; McClimon, J Brandon; Rose, Franck; Carpick, Robert W

    2014-12-16

    Near-edge X-ray absorption fine structure (NEXAFS) spectroscopy is a powerful technique for characterizing the composition and bonding state of nanoscale materials and the top few nanometers of bulk and thin film specimens. When coupled with imaging methods like photoemission electron microscopy, it enables chemical imaging of materials with nanometer-scale lateral spatial resolution. However, analysis of NEXAFS spectra is often performed under the assumption of structural and compositional homogeneity within the nanometer-scale depth probed by this technique. This assumption can introduce large errors when analyzing the vast majority of solid surfaces due to the presence of complex surface and near-surface structures such as oxides and contamination layers. An analytical methodology is presented for removing the contribution of these nanoscale overlayers from NEXAFS spectra of two-layered systems to provide a corrected photoabsorption spectrum of the substrate. This method relies on the subtraction of the NEXAFS spectrum of the overlayer adsorbed on a reference surface from the spectrum of the two-layer system under investigation, where the thickness of the overlayer is independently determined by X-ray photoelectron spectroscopy (XPS). This approach is applied to NEXAFS data acquired for one of the most challenging cases: air-exposed hard carbon-based materials with adventitious carbon contamination from ambient exposure. The contribution of the adventitious carbon was removed from the as-acquired spectra of ultrananocrystalline diamond (UNCD) and hydrogenated amorphous carbon (a-C:H) to determine the intrinsic photoabsorption NEXAFS spectra of these materials. The method alters the calculated fraction of sp(2)-hybridized carbon from 5 to 20% and reveals that the adventitious contamination can be described as a layer containing carbon and oxygen ([O]/[C] = 0.11 ± 0.02) with a thickness of 0.6 ± 0.2 nm and a fraction of sp(2)-bonded carbon of 0.19 ± 0.03. This

  4. Ectopic expression of class 1 KNOX genes induce adventitious shoot regeneration and alter growth and development of tobacco (Nicotiana tabacum L) and European plum (Prunus domestica L).

    PubMed

    Srinivasan, C; Liu, Zongrang; Scorza, Ralph

    2011-04-01

    Transgenic plants of tobacco (Nicotiana tabacum L) and European plum (Prunus domestica L) were produced by transforming with the apple class 1 KNOX genes (MdKN1 and MdKN2) or corn KNOX1 gene. Transgenic tobacco plants were regenerated in vitro from transformed leaf discs cultured in a medium lacking cytokinin. Ectopic expression of KNOX genes retarded shoot growth by suppressing elongation of internodes in transgenic tobacco plants. Expression of each of the three KNOX1 genes induced malformation and extensive lobbing in tobacco leaves. In situ regeneration of adventitious shoots was observed from leaves and roots of transgenic tobacco plants expressing each of the three KNOX genes. In vitro culture of leaf explants and internode sections excised from in vitro grown MdKN1 expressing tobacco shoots regenerated adventitious shoots on MS (Murashige and Skoog 1962) basal medium in the absence of exogenous cytokinin. Transgenic plum plants that expressed the MdKN2 or corn KNOX1 gene grew normally but MdKN1 caused a significant reduction in plant height, leaf shape and size and produced malformed curly leaves. A high frequency of adventitious shoot regeneration (96%) was observed in cultures of leaf explants excised from corn KNOX1-expressing transgenic plum shoots. In contrast to KNOX1-expressing tobacco, leaf and internode explants of corn KNOX1-expressing plum required synthetic cytokinin (thidiazuron) in the culture medium to induce adventitious shoot regeneration. The induction of high-frequency regeneration of adventitious shoots in vitro from leaves and stem internodal sections of plum through the ectopic expression of a KNOX1 gene is the first such report for a woody perennial fruit trees.

  5. Production of biomass and bioactive compounds from adventitious roots by optimization of culturing conditions of Eurycoma longifolia in balloon-type bubble bioreactor system.

    PubMed

    Lulu, Tao; Park, So-Young; Ibrahim, Rusli; Paek, Kee-Yoeup

    2015-06-01

    The present study aimed to optimize the conditions for the production of adventitious roots from Eurycoma longifolia Jack, an important medicinal woody plant, in bioreactor culture. The effects of the type and concentration of auxin on root growth were studied, as well as the effects of the NH4(+):NO3(-) ratio on adventitious root growth and the production of phenolics and flavonoids. Approximately 5 g L(-1) fresh weight of adventitious roots was inoculated into a 3 L balloon-type bubble bioreactor, which contained 2 L 3/4 MS medium supplemented with 30 g L(-1) sucrose and cultures were maintained in the dark for 7 weeks at 24 ± 1°C. Higher concentrations of IBA (7.0 and 9.0 mg L(-1)) and NAA (5.0 mg L(-1)) enhanced the biomass and accumulation of total phenolics and flavonoids. The adventitious roots were thin, numerous, and elongated in 3/4 MS medium supplemented with 5.0 and 7.0 mg L(-1) IBA, whereas the lateral roots were shorter and thicker with 5.0 mg L(-1) NAA compared with IBA treatment. The optimum biomasses of 50.22 g L(-1) fresh weight and 4.60 g L(-1) dry weight were obtained with an NH4(+):NO3(-) ratio of 15:30. High phenolic and flavonoid productions (38.59 and 11.27 mg L(-1) medium, respectively) were also obtained with a ratio of 15:30. Analysis of the 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging activity indicated higher antioxidant activity with an NH4(+):NO3(-) ratio of 30:15. These results suggest that balloon-type bubble bioreactor cultures are suitable for the large-scale commercial production of E. longifolia adventitious roots which contain high yield of bioactive compounds.

  6. Arterial Wall Perfusion Measured with Photon Counting Spectral X-ray CT

    PubMed Central

    Jorgensen, Steven M.; Korinek, Mark J.; Vercnocke, Andrew J.; Anderson, Jill L.; Halaweish, Ahmed; Leng, Shuai; McCollough, Cynthia H.; Ritman, Erik L.

    2016-01-01

    Early atherosclerosis changes perfusion of the arterial wall due to localized proliferation of the vasa vasorum. When contrast agent passes through the artery, some enters the vasa vasorum and increases radiopacity of the arterial wall. Technical challenges to detecting changes in vasa vasorum density include the thin arterial wall, partial volume averaging at the arterial lumen/wall interface and calcification within the wall. We used a photon-counting spectral CT scanner to study carotid arteries of anesthetized pigs and micro-CT of these arteries to quantify vasa vasorum density. The left carotid artery wall was injected with autologous blood to stimulate vasa vasorum angiogenesis. The scans were performed at 25–120 keV; the tube-current-time product was 550 mAs. A 60 mL bolus of iodine contrast agent was injected into the femoral vein at 5mL/s. Two seconds post injection, an axial scan was acquired at every 3 s over 60 s (i.e., 20 time points). Each time point acquired 28 contiguous transaxial slices with reconstructed voxels 0.16 × 0.16 × 1 mm3. Regions-of-interest in the outer 2/3 of the arterial wall and in the middle 2/3 of the lumen were drawn and their enhancements plotted versus time. Lumenal CT values peaked several seconds after injection and then returned towards baseline. Arterial wall CT values peaked concurrent to the lumen. The peak arterial wall enhancement in the left carotid arterial wall correlated with increased vasa vasorum density observed in micro-CT images of the isolated arteries. PMID:27807391

  7. Ethylene and adventitious root formation in hypocotyl segments of etiolated mung-bean (Vigna radiata (L.) Wilczek) seedlings.

    PubMed

    Batten, D J; Mullins, M G

    1978-01-01

    Rooting responses and ethylene production by hypocotyl cuttings from etiolated mung-bean seedlings treated with the auxins α-naphthaleneacetic acid, γ-(indole-3)-n-butyric acid (IBA) and 2,4,5-trichloro-phenoxypropionic acid were determined. There was no relationship between the abilities of the auxins to induce root formation and their capacities for inducing ethylene production. Studies with mixtures of 3-indoleacetic acid, a poor stimulator of rooting but an effective inducer of ethylene production, and IBA, an effective rooting stimulator but a poor inducer of ethylene production, exposure of cuttings to ethylene or (2-chloroethyl) phosphonic acid (Ethephon), hypobaric storage (150 mb) of treated cuttings, and exposure of auxin-treated cuttings to 7% CO2 also indicated that ethylene is not directly involved in initiation of adventitious roots in this plant material. PMID:24414045

  8. Direct adventitious shoot bud formation on hypocotyls explants in Millettia pinnata (L.) Panigrahi- a biodiesel producing medicinal tree species.

    PubMed

    Nagar, Durga Singh; Jha, Suman Kumar; Jani, Jigar

    2015-04-01

    A reproducible protocol developed for in vitro regeneration of Milletia pinnata using hypocotyl segments. Multiple shoots were induced from hypocotyl explants through direct adventitious shoot bud regeneration. The proximal end of hypocotyls was responsive for shoot bud induction. Silver nitrate and adenine sulphate had a positive effect on shoot bud induction and elongation. The maximum response and number of shoot bud produced in media supplemented with 8.88 μM BAP with 108.6 μM adenine sulphate and 11.84 μM silver nitrate. Elongated shoots were harvested and successful rooting of microshoots achieved on MS media supplemented with 9.84 μM IBA, with 81.1 % rooting. Remaining shoot buds sub-cultured for further multiplication and elongation. Each subculture produced eight to nine elongated microshoots up to four subcultures. The rooted microshoots were successfully hardened and transferred to field. PMID:25964721

  9. Activity and isoforms of peroxidases, lignin and anatomy, during adventitious rooting in cuttings of Ebenus cretica L.

    PubMed

    Syros, Thomas; Yupsanis, Traianos; Zafiriadis, Helias; Economou, Athanasios

    2004-01-01

    Adventitious rooting of Ebenus cretica cuttings was studied in order to examine a) the rooting ability of different genotypes in relation to electrophoretic patterns of peroxidases. b) the activity and electrophoretic patterns of soluble and wall ionically bound peroxidases, the lignin content and anatomical changes in the control and IBA treated cuttings of and genotypes in the course of adventitious root formation. In addition, a fraction of soluble cationic peroxidases was separated by gel filtration chromatography from the total soluble peroxidases of a genotype. No rooting occurred in cuttings without IBA-treatment. In both genotypes, electrophoretic patterns of soluble anionic peroxidases revealed two common peroxidase isoforms, while a fast-migrating anionic peroxidase isoform (A3) appeared only in genotypes. Both genotypes showed similar patterns of soluble, as well as wall ionically bound cationic peroxidase isoforms. The number of isoforms was unchanged during the rooting process (induction, initiation and expression phase) but an increase in peroxidase activity (initiation phase) followed by decrease has been found in IBA-treated cuttings. During initiation phase the lignin content was almost similar to that on day 0 in genotype while it was reduced at by about 50% in genotype at the respective time. Microscopic observations revealed anatomical differences between genotypes. According to this study, the and genotypes display differences in anatomy, lignin content, activity of soluble peroxidases and the electrophoretic patterns of soluble anionic peroxidase isoforms. The A3-anionic peroxidase isoform could be used as biochemical marker to distinguish and genotypes of E. cretica and seems to be correlated to lignin synthesis in rooting process. PMID:15002666

  10. Adventitious Arsenate Reductase Activity of the Catalytic Domain of the Human Cdc25B and Cdc25C Phosphatases†

    PubMed Central

    Bhattacharjee, Hiranmoy; Sheng, Ju; Ajees, A. Abdul; Mukhopadhyay, Rita; Rosen, Barry P.

    2013-01-01

    A number of eukaryotic enzymes that function as arsenate reductases are homologues of the catalytic domain of the human Cdc25 phosphatase. For example, the Leishmania major enzyme LmACR2 is both a phosphatase and an arsenate reductase, and its structure bears similarity to the structure of the catalytic domain of human Cdc25 phosphatase. These reductases contain an active site C-X5-R signature motif, where C is the catalytic cysteine, the five X residues form a phosphate binding loop, and R is a highly conserved arginine, which is also present in human Cdc25 phosphatases. We therefore investigated the possibility that the three human Cdc25 isoforms might have adventitious arsenate reductase activity. The sequences for the catalytic domains of Cdc25A, -B, and -C were cloned individually into a prokaryotic expression vector, and their gene products were purified from a bacterial host using nickel affinity chromatography. While each of the three Cdc25 catalytic domains exhibited phosphatase activity, arsenate reductase activity was observed only with Cdc25B and -C. These two enzymes reduced inorganic arsenate but not methylated pentavalent arsenicals. Alteration of either the cysteine and arginine residues of the Cys-X5-Arg motif led to the loss of both reductase and phosphatase activities. Our observations suggest that Cdc25B and -C may adventitiously reduce arsenate to the more toxic arsenite and may also provide a framework for identifying other human protein tyrosine phosphatases containing the active site Cys-X5-Arg loop that might moonlight as arsenate reductases. PMID:20025242

  11. Effect of nitrogen source on biomass and bioactive compound production in submerged cultures of Eleutherococcus koreanum Nakai adventitious roots.

    PubMed

    Lee, Eun-Jung; Paek, Kee-Yoeup

    2012-01-01

    Ammonium to nitrate ratios of 0:30, 5:25, 10:20, 15:15, 20:10, 25:5, and 30:0 mM were tested to determine the optimal NH(4)(+) :NO(3)(-) ratio for improving biomass and bioactive compound production in Eleutherococcus koreanum Nakai adventitious roots using 3-L bulb-type bubble bioreactors. A high ammonium nitrogen ratio had a negative effect on root growth, and the highest fresh and dry weights were obtained when NH(4)(+):NO(3)(-) ratios were 5:25 and 10:20 (mM) after 5 weeks of culture. Although the total production of eleutherosides B and E was slightly higher at the 10:20 ratio than at the 5:25 ratio (NH(4)(+):NO(3)(-)), we proposed that the optimal NH(4)(+):NO(3)(-) ratio was 5:25 mM. This ratio achieved both the highest total production of five target bioactive compounds (eleutherosides B and E, chlorogenic acid, total phenolics, and flavonoids) and the highest root biomass. Furthermore, increasing NH(4)(+):NO(3)(-) ratios to 10:20 decreased pH in the medium, interrupted the absorption of essential minerals from the culture medium, and resulted in low biomass and increased relative oxidative stress levels, which were evaluated by determining 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Therefore, nitrate rather than ammonium nitrogen was more essential not for only biomass production but also for bioactive compound production in E. koreanum adventitious root cultures. The optimal nitrogen source ratio produced 5.63 g L(-1) of biomass and 24.41 mg of the five total bioactive compounds per gram of biomass (dry weight basis). The development of such in vitro culture technology will benefit the pilot-scale production of E. koreanum-based bioactive compounds for commercialization.

  12. Modeling gene flow distribution within conventional fields and development of a simplified sampling method to quantify adventitious GM contents in maize.

    PubMed

    Melé, Enric; Nadal, Anna; Messeguer, Joaquima; Melé-Messeguer, Marina; Palaudelmàs, Montserrat; Peñas, Gisela; Piferrer, Xavier; Capellades, Gemma; Serra, Joan; Pla, Maria

    2015-01-01

    Genetically modified (GM) crops have been commercially grown for two decades. GM maize is one of 3 species with the highest acreage and specific events. Many countries established a mandatory labeling of products containing GM material, with thresholds for adventitious presence, to support consumers' freedom of choice. In consequence, coexistence systems need to be introduced to facilitate commercial culture of GM and non-GM crops in the same agricultural area. On modeling adventitious GM cross-pollination distribution within maize fields, we deduced a simple equation to estimate overall GM contents (%GM) of conventional fields, irrespective of its shape and size, and with no previous information on possible GM pollen donor fields. A sampling strategy was designed and experimentally validated in 19 agricultural fields. With 9 samples, %GM quantification requires just one analytical GM determination while identification of the pollen source needs 9 additional analyses. A decision support tool is provided. PMID:26596213

  13. A gastric artery aneurysm complicated by a dissection of gastric and hepatic arteries: possible role of adventitial inflammation and disruption of internal elastic lamina in splanchnic artery dissection.

    PubMed

    Kobayashi, Masahiko; Mellen, Paul F

    2008-06-01

    A 77-year-old woman was found deceased at home. An autopsy examination revealed a hemoperitoneum due to a ruptured false aneurysm of a branch of the left gastric artery. A long dissection extending from the aneurysm involved splanchnic arteries including the left gastric, common hepatic, right and left branches of proper hepatic, and intrahepatic arteries. An intimal tear was identified in the common hepatic artery. Neutrophils infiltrating in the adventitia may have been reactive and may have triggered the adventitial rupture of aneurysm or development of the dissection. Disruption of the internal elastic lamina, which has been proposed to cause dissection of intracranial arteries, was seen in the dissected arteries. Little is currently known about aneurysms or dissections of splanchnic arteries; however, observation of adventitial inflammation and internal elastic lamina may help disclose the etiology and pathogenesis. PMID:18520493

  14. Modeling gene flow distribution within conventional fields and development of a simplified sampling method to quantify adventitious GM contents in maize

    PubMed Central

    Melé, Enric; Nadal, Anna; Messeguer, Joaquima; Melé-Messeguer, Marina; Palaudelmàs, Montserrat; Peñas, Gisela; Piferrer, Xavier; Capellades, Gemma; Serra, Joan; Pla, Maria

    2015-01-01

    Genetically modified (GM) crops have been commercially grown for two decades. GM maize is one of 3 species with the highest acreage and specific events. Many countries established a mandatory labeling of products containing GM material, with thresholds for adventitious presence, to support consumers’ freedom of choice. In consequence, coexistence systems need to be introduced to facilitate commercial culture of GM and non-GM crops in the same agricultural area. On modeling adventitious GM cross-pollination distribution within maize fields, we deduced a simple equation to estimate overall GM contents (%GM) of conventional fields, irrespective of its shape and size, and with no previous information on possible GM pollen donor fields. A sampling strategy was designed and experimentally validated in 19 agricultural fields. With 9 samples, %GM quantification requires just one analytical GM determination while identification of the pollen source needs 9 additional analyses. A decision support tool is provided. PMID:26596213

  15. PDA/FDA Adventitious Agents and Novel Cell Substrates: Emerging Technologies and New Challenges, Nov. 3-4, 2011, Rockville, MD.

    PubMed

    Khan, Arifa S; Lubiniecki, Anthony; King, Kathryn E

    2012-01-01

    In response to the challenges of addressing potential safety concerns related to the use of novel cell substrates, the Parenteral Drug Association (PDA) and the U.S. Food and Drug Administration (FDA) co-sponsored a conference on November 3-4, 2011 in Rockville, Maryland, USA. The following were discussed: the current adventitious agent testing methods and the use of emerging nucleic acid-based technologies for broad detection of known and novel viruses; issues associated with manufacture of biologicals in novel cell substrates such as insect cell lines and plants, with specific focus on potential safety concerns related to endogenous retroviral elements; and raw material controls and strategies to mitigate risk of adventitious virus contamination during product manufacture.

  16. Identification of genes involved in indole-3-butyric acid-induced adventitious root formation in nodal cuttings of Camellia sinensis (L.) by suppression subtractive hybridization.

    PubMed

    Wei, Kang; Wang, Liyuan; Cheng, Hao; Zhang, Chengcai; Ma, Chunlei; Zhang, Liqun; Gong, Wuyun; Wu, Liyun

    2013-02-10

    The plant hormone auxin plays a key role in adventitious rooting. To increase our understanding of genes involved in adventitious root formation, we identified transcripts differentially expressed in single nodal cuttings of Camellia sinensis treated with or without indole-3-butyric acid (IBA) by suppressive subtractive hybridization (SSH). A total of 77 differentially expressed transcripts, including 70 up-regulated and 7 down-regulated sequences, were identified in tea cuttings under IBA treatment. Seven candidate transcripts were selected and analyzed for their response to IBA, and IAA by real time RT-PCR. All these transcripts were up regulated by at least two folds one day after IBA treatment. Meanwhile, IAA showed less positive effects on the expression of candidate transcripts. The full-length cDNA of a F-box/kelch gene was also isolated and found to be similar to a group of At1g23390 like genes. These unigenes provided a new source for mining genes related to adventitious root formation, which facilitate our understanding of relative fundamental metabolism.

  17. Identification of genes involved in indole-3-butyric acid-induced adventitious root formation in nodal cuttings of Camellia sinensis (L.) by suppression subtractive hybridization.

    PubMed

    Wei, Kang; Wang, Liyuan; Cheng, Hao; Zhang, Chengcai; Ma, Chunlei; Zhang, Liqun; Gong, Wuyun; Wu, Liyun

    2013-02-10

    The plant hormone auxin plays a key role in adventitious rooting. To increase our understanding of genes involved in adventitious root formation, we identified transcripts differentially expressed in single nodal cuttings of Camellia sinensis treated with or without indole-3-butyric acid (IBA) by suppressive subtractive hybridization (SSH). A total of 77 differentially expressed transcripts, including 70 up-regulated and 7 down-regulated sequences, were identified in tea cuttings under IBA treatment. Seven candidate transcripts were selected and analyzed for their response to IBA, and IAA by real time RT-PCR. All these transcripts were up regulated by at least two folds one day after IBA treatment. Meanwhile, IAA showed less positive effects on the expression of candidate transcripts. The full-length cDNA of a F-box/kelch gene was also isolated and found to be similar to a group of At1g23390 like genes. These unigenes provided a new source for mining genes related to adventitious root formation, which facilitate our understanding of relative fundamental metabolism. PMID:23201417

  18. Molecular cloning, characterization and expression analysis of the SAMS gene during adventitious root development in IBA-induced tetraploid black locust.

    PubMed

    Quan, Jine; Zhang, Sheng; Zhang, Chunxia; Meng, Sen; Zhao, Zhong; Xu, Xuexuan

    2014-01-01

    S-Adenosylmethionine synthetase (SAMS) catalyzes the synthesis of S-adenosylmethionine (SAM), a precursor for ethylene and polyamine biosynthesis. Here, we report the isolation of the 1498 bp full-length cDNA sequence encoding tetraploid black locust (Robinia pseudoacacia L.) SAMS (TrbSAMS), which contains an open reading frame of 1179 bp encoding 392 amino acids. The amino acid sequence of TrbSAMS has more than 94% sequence identity to SAMSs from other plants, with a closer phylogenetic relationship to SAMSs from legumes than to SAMS from other plants. The TrbSAMS monomer consists of N-terminal, central, and C-terminal domains. Subcellular localization analysis revealed that the TrbSAMS protein localizes mainly to in the cell membrane and cytoplasm of onion epidermal cells and Arabidopsis mesophyll cell protoplasts. Indole-3-butyric acid (IBA)-treated cuttings showed higher levels of TrbSAMS transcript than untreated control cuttings during root primordium and adventitious root formation. TrbSAMS and its downstream genes showed differential expression in shoots, leaves, bark, and roots, with the highest expression observed in bark. IBA-treated cuttings also showed higher SAMS activity than control cuttings during root primordium and adventitious root formation. These results indicate that TrbSAMS might play an important role in the regulation of IBA-induced adventitious root development in tetraploid black locust cuttings. PMID:25285660

  19. Nuclear localization and interaction of RolB with plant 14-3-3 proteins correlates with induction of adventitious roots by the oncogene rolB.

    PubMed

    Moriuchi, Hiroshi; Okamoto, Chiho; Nishihama, Ryuichi; Yamashita, Ichiro; Machida, Yasunori; Tanaka, Nobukazu

    2004-04-01

    The rooting-locus gene B (rolB) on the T-DNA of the root-inducing (Ri) plasmid in Agrobacterium rhizogenes is responsible for the induction of transformed adventitious roots, although the root induction mechanism is unknown. We report here that the RolB protein of pRi1724 (1724RolB) is associated with Nicotianatabacum14-3-3-like protein omegaII (Nt14-3-3 omegaII) in tobacco bright yellow (BY)-2 cells. Nt14-3-3 omegaII directly interacts with 1724RolB protein. Green fluorescent protein (GFP)-fused 1724RolB is localized to the nucleus. GFP-fused mutant 1724RolB proteins having a deletion or amino acid substitution are unable to interact with Nt14-3-3 omegaII and also show impaired nuclear localization. Moreover, these 1724RolB mutants show decreased capacity for adventitious root induction. These results suggest that adventitious root induction by 1724RolB protein correlates with its interaction with Nt14-3-3 omegaII and the nuclear localization of 1724RolB protein. PMID:15078329

  20. Rhabdovirus-like endogenous viral elements in the genome of Spodoptera frugiperda insect cells are actively transcribed: Implications for adventitious virus detection.

    PubMed

    Geisler, Christoph; Jarvis, Donald L

    2016-07-01

    Spodoptera frugiperda (Sf) cell lines are used to produce several biologicals for human and veterinary use. Recently, it was discovered that all tested Sf cell lines are persistently infected with Sf-rhabdovirus, a novel rhabdovirus. As part of an effort to search for other adventitious viruses, we searched the Sf cell genome and transcriptome for sequences related to Sf-rhabdovirus. To our surprise, we found intact Sf-rhabdovirus N- and P-like ORFs, and partial Sf-rhabdovirus G- and L-like ORFs. The transcribed and genomic sequences matched, indicating the transcripts were derived from the genomic sequences. These appear to be endogenous viral elements (EVEs), which result from the integration of partial viral genetic material into the host cell genome. It is theoretically impossible for the Sf-rhabdovirus-like EVEs to produce infectious virus particles as 1) they are disseminated across 4 genomic loci, 2) the G and L ORFs are incomplete, and 3) the M ORF is missing. Our finding of transcribed virus-like sequences in Sf cells underscores that MPS-based searches for adventitious viruses in cell substrates used to manufacture biologics should take into account both genomic and transcribed sequences to facilitate the identification of transcribed EVE's, and to avoid false positive detection of replication-competent adventitious viruses.

  1. Molecular Cloning, Characterization and Expression Analysis of the SAMS Gene during Adventitious Root Development in IBA-Induced Tetraploid Black Locust

    PubMed Central

    Quan, Jine; Zhang, Sheng; Zhang, Chunxia; Meng, Sen; Zhao, Zhong; Xu, Xuexuan

    2014-01-01

    S-Adenosylmethionine synthetase (SAMS) catalyzes the synthesis of S-adenosylmethionine (SAM), a precursor for ethylene and polyamine biosynthesis. Here, we report the isolation of the 1498 bp full-length cDNA sequence encoding tetraploid black locust (Robinia pseudoacacia L.) SAMS (TrbSAMS), which contains an open reading frame of 1179 bp encoding 392 amino acids. The amino acid sequence of TrbSAMS has more than 94% sequence identity to SAMSs from other plants, with a closer phylogenetic relationship to SAMSs from legumes than to SAMS from other plants. The TrbSAMS monomer consists of N-terminal, central, and C-terminal domains. Subcellular localization analysis revealed that the TrbSAMS protein localizes mainly to in the cell membrane and cytoplasm of onion epidermal cells and Arabidopsis mesophyll cell protoplasts. Indole-3-butyric acid (IBA)-treated cuttings showed higher levels of TrbSAMS transcript than untreated control cuttings during root primordium and adventitious root formation. TrbSAMS and its downstream genes showed differential expression in shoots, leaves, bark, and roots, with the highest expression observed in bark. IBA-treated cuttings also showed higher SAMS activity than control cuttings during root primordium and adventitious root formation. These results indicate that TrbSAMS might play an important role in the regulation of IBA-induced adventitious root development in tetraploid black locust cuttings. PMID:25285660

  2. Assessing the three-dimensional collagen network in soft tissues using contrast agents and high resolution micro-CT: Application to porcine iliac veins.

    PubMed

    Nierenberger, Mathieu; Rémond, Yves; Ahzi, Saïd; Choquet, Philippe

    2015-07-01

    The assessment of the three-dimensional architecture of collagen fibers inside vessel walls constitutes one of the bases for building structural models for the description of the mechanical behavior of these tissues. Multiphoton microscopy allows for such observations, but is limited to volumes of around a thousand of microns. In the present work, we propose to observe the collagenous network of vascular tissues using micro-CT. To get a contrast, three staining solutions (phosphotungstic acid, phosphomolybdic acid and iodine potassium iodide) were tested. Two of these stains were showed to lead to similar results and to a satisfactory contrast within the tissue. A detailed observation of a small porcine iliac vein sample allowed assessing the collagen fibers orientations within the medial and adventitial layers of the vein. The vasa vasorum network, which is present inside the adventitia of the vein, was also observed. Finally, the demonstrated micro-CT staining technique for the three-dimensional observation of thin soft tissues samples, like vein walls, contributes to the assessment of their structure at different scales while keeping a global overview of the tissue. PMID:26033495

  3. Focus on the "unstable" carotid plaque: detection of intraplaque angiogenesis with contrast ultrasound. Present state and future perspectives.

    PubMed

    Giannoni, Maria Fabrizia; Vicenzini, Edoardo

    2009-04-01

    Over the past 20 years, conventional ultrasonography has identified features of the "unstable" carotid plaque. Histological studies have recognized that plaque inflammation and neoangiogenesis play a pivotal role in the developing of the vulnerable plaque. Hence, the growing interest on the biological activities of atherosclerotic lesions leading to cerebrovascular events. The presence of adventitial vasa vasorum and the occurrence of plaque vascularization have been considered as predictors of unstable lesions in cerebrovascular and/or cardiovascular patients. The advent of ultrasound contrast agents has represented a fundamental step in the up-to-date functional evaluation in several fields with minimally invasive procedures. Contrast specific ultrasound modalities are currently used with excellent results in oncology, in cardiology and in vascular diseases. Contrast carotid ultrasound is an emerging imaging technique, able to depict in vivo new functional information on plaque activity and vascularization that may add further new data on the actual condition and future cerebrovascular risk. Further studies will provide a better clarification of the degree of neo-angiogenesis. A future strategy could be represented by the monitoring of plaque neoangiogenesis in order to detect the possible pharmacological effects on plaque remodeling.

  4. Evidence for vocal learning in juvenile male killer whales, Orcinus orca, from an adventitious cross-socializing experiment.

    PubMed

    Crance, Jessica L; Bowles, Ann E; Garver, Alan

    2014-04-15

    Killer whales (Orcinus orca) are thought to learn their vocal dialect. Dispersal in the species is rare, but effects of shifts in social association on the dialect can be studied under controlled conditions. Individual call repertoires and social association were measured in three adult female killer whales and three males (two juveniles and an adult) during two periods, 2001-2003 and 2005-2006. Three distinct dialect repertoires were represented among the subjects. An adventitious experiment in social change resulted from the birth of a calf and the transfer of two non-focal subjects in 2004. Across the two periods, 1691 calls were collected, categorized and attributed to individuals. Repertoire overlap for each subject dyad was compared with an index of association. During 2005-2006, the two juvenile males increased association with the unrelated adult male. By the end of the period, both had begun producing novel calls and call features characteristic of his repertoire. However, there was little or no reciprocal change and the adult females did not acquire his calls. Repertoire overlap and association were significantly correlated in the first period. In the second, median association time and repertoire similarity increased, but the relationship was only marginally significant. The results provided evidence that juvenile male killer whales are capable of learning new call types, possibly stimulated by a change in social association. The pattern of learning was consistent with a selective convergence of male repertoires. PMID:24744421

  5. In vitro adventitious shoot regeneration via indirect organogenesis from petiole explants of Cassia angustifolia Vahl.-a potential medicinal plant.

    PubMed

    Siddique, Iram; Anis, M; Aref, I M

    2010-11-01

    An effective protocol was developed for in vitro regeneration of the Cassia angustifolia via indirect organogenesis from petiole explants excised from 21-day-old axenic seedlings. Organogenic callus were induced on Murashige and Skoog (MS) medium supplemented with 5.0 µM 2,4-dichlorophenoxy acetic acid and 2.5 µM thidiazuron (TDZ). Adventitious shoot regeneration was achieved on MS medium supplemented with 5.0 µM TDZ as it induced 8.5 ± 0.98 shoots in 85% cultures. The number of shoots and shoot length was significantly enhanced when cultures were subcultured on auxin-cytokinin-containing medium. The highest number of shoots (12.5 ± 1.10) and shoot length (4.3 ± 0.20 cm) was recorded on MS medium supplemented with 5.0 µM TDZ and 1.5 µM indole-3-acetic acid. Regenerated shoots were rooted best on MS medium supplemented with 10.0 µM indole-3-butyric acid followed by their transfer to liquid MS filter paper bridge medium. The plants were successfully hardened off in sterile soilrite followed by their establishment in garden soil with 70% survival rate. The plants showed normal morphological characteristics similar to the field grown plants. PMID:20461494

  6. Effect of Naphthalene Acetic Acid on Adventitious Root Development and Associated Physiological Changes in Stem Cutting of Hemarthria compressa

    PubMed Central

    Zhang, Xin-Quan; Yang, Wen-Yu; Wan, Yan; Ma, Ying-Mei; Zhu, Yong-Qun; Peng, Yan; Huang, Lin-Kai

    2014-01-01

    In order to find a way to induce rooting on cuttings of Hemarthria compressa cv. Ya’an under controlled conditions, a project was carried out to study the effect of naphthalene acetic acid (NAA) on rooting in stem cuttings and related physiological changes during the rooting process of Hemarthria compressa. The cuttings were treated with five concentrations of NAA (0, 100, 200 300, 400 mg/l) at three soaking durations (10, 20, 30 minutes), and cuttings without treatment were considered as control. Samples were planted immediately into pots after treatment. IAA-oxidase (IAAO) activity, peroxidase (POD) activity and polyphenol oxidase (PPO) activity were determined after planting. Results showed that NAA had positive effect on rooting at the concentration of 200 mg/l compared to other concentrations at 30 days after planting (DAP). Among the three soaking durations, 20 minutes (min) of 200 mg/l NAA resulted in higher percentages of rooting, larger numbers of adventitious roots and heavier root dry weight per cutting. The lowest IAAO activity was obtained when soaked at 200 mg/l NAA for 20 min soaking duration. This was consistent with the best rooting ability, indicating that the lower IAAO activity, the higher POD activity and PPO activity could be used as an indicator of better rooting ability for whip grass cuttings and might serve as a good marker for rooting ability in cuttings. PMID:24595064

  7. Evidence for vocal learning in juvenile male killer whales, Orcinus orca, from an adventitious cross-socializing experiment.

    PubMed

    Crance, Jessica L; Bowles, Ann E; Garver, Alan

    2014-04-15

    Killer whales (Orcinus orca) are thought to learn their vocal dialect. Dispersal in the species is rare, but effects of shifts in social association on the dialect can be studied under controlled conditions. Individual call repertoires and social association were measured in three adult female killer whales and three males (two juveniles and an adult) during two periods, 2001-2003 and 2005-2006. Three distinct dialect repertoires were represented among the subjects. An adventitious experiment in social change resulted from the birth of a calf and the transfer of two non-focal subjects in 2004. Across the two periods, 1691 calls were collected, categorized and attributed to individuals. Repertoire overlap for each subject dyad was compared with an index of association. During 2005-2006, the two juvenile males increased association with the unrelated adult male. By the end of the period, both had begun producing novel calls and call features characteristic of his repertoire. However, there was little or no reciprocal change and the adult females did not acquire his calls. Repertoire overlap and association were significantly correlated in the first period. In the second, median association time and repertoire similarity increased, but the relationship was only marginally significant. The results provided evidence that juvenile male killer whales are capable of learning new call types, possibly stimulated by a change in social association. The pattern of learning was consistent with a selective convergence of male repertoires.

  8. Oxygen deficiency and salinity affect cell-specific ion concentrations in adventitious roots of barley (Hordeum vulgare).

    PubMed

    Kotula, Lukasz; Clode, Peta L; Striker, Gustavo G; Pedersen, Ole; Läuchli, André; Shabala, Sergey; Colmer, Timothy D

    2015-12-01

    Oxygen deficiency associated with soil waterlogging adversely impacts root respiration and nutrient acquisition. We investigated the effects of O2 deficiency and salinity (100 mM NaCl) on radial O2 concentrations and cell-specific ion distributions in adventitious roots of barley (Hordeum vulgare). Microelectrode profiling measured O2 concentrations across roots in aerated, aerated saline, stagnant or stagnant saline media. X-ray microanalysis at two positions behind the apex determined the cell-specific elemental concentrations of potassium (K), sodium (Na) and chloride (Cl) across roots. Severe O2 deficiency occurred in the stele and apical regions of roots in stagnant solutions. O2 deficiency in the stele reduced the concentrations of K, Na and Cl in the pericycle and xylem parenchyma cells at the subapical region. Near the root apex, Na declined across the cortex in roots from the aerated saline solution but was relatively high in all cell types in roots from the stagnant saline solution. Oxygen deficiency has a substantial impact on cellular ion concentrations in roots. Both pericycle and xylem parenchyma cells are involved in energy-dependent K loading into the xylem and in controlling radial Na and Cl transport. At root tips, accumulation of Na in the outer cell layers likely contributed to reduction of Na in inner cells of the tips.

  9. A Co-Opted Hormonal Cascade Activates Dormant Adventitious Root Primordia upon Flooding in Solanum dulcamara1[OPEN

    PubMed Central

    Dawood, Thikra; Kensche, Philip R.; Cristescu, Simona M.; Mariani, Celestina

    2016-01-01

    Soil flooding is a common stress factor affecting plants. To sustain root function in the hypoxic environment, flooding-tolerant plants may form new, aerenchymatous adventitious roots (ARs), originating from preformed, dormant primordia on the stem. We investigated the signaling pathway behind AR primordium reactivation in the dicot species Solanum dulcamara. Transcriptome analysis indicated that flooding imposes a state of quiescence on the stem tissue, while increasing cellular activity in the AR primordia. Flooding led to ethylene accumulation in the lower stem region and subsequently to a drop in abscisic acid (ABA) level in both stem and AR primordia tissue. Whereas ABA treatment prevented activation of AR primordia by flooding, inhibition of ABA synthesis was sufficient to activate them in absence of flooding. Together, this reveals that there is a highly tissue-specific response to reduced ABA levels. The central role for ABA in the response differentiates the pathway identified here from the AR emergence pathway known from rice (Oryza sativa). Flooding and ethylene treatment also induced expression of the polar auxin transporter PIN2, and silencing of this gene or chemical inhibition of auxin transport inhibited primordium activation, even though ABA levels were reduced. Auxin treatment, however, was not sufficient for AR emergence, indicating that the auxin pathway acts in parallel with the requirement for ABA reduction. In conclusion, adaptation of S. dulcamara to wet habitats involved co-option of a hormonal signaling cascade well known to regulate shoot growth responses, to direct a root developmental program upon soil flooding. PMID:26850278

  10. Transcriptomic analysis reveals ethylene as stimulator and auxin as regulator of adventitious root formation in petunia cuttings

    PubMed Central

    Druege, Uwe; Franken, Philipp; Lischewski, Sandra; Ahkami, Amir H.; Zerche, Siegfried; Hause, Bettina; Hajirezaei, Mohammad R.

    2014-01-01

    Adventitious root (AR) formation in the stem base (SB) of cuttings is the basis for propagation of many plant species and petunia is used as model to study this developmental process. Following AR formation from 2 to 192 hours post-excision (hpe) of cuttings, transcriptome analysis by microarray revealed a change of the character of the rooting zone from SB to root identity. The greatest shift in the number of differentially expressed genes was observed between 24 and 72 hpe, when the categories storage, mineral nutrient acquisition, anti-oxidative and secondary metabolism, and biotic stimuli showed a notable high number of induced genes. Analyses of phytohormone-related genes disclosed multifaceted changes of the auxin transport system, auxin conjugation and the auxin signal perception machinery indicating a reduction in auxin sensitivity and phase-specific responses of particular auxin-regulated genes. Genes involved in ethylene biosynthesis and action showed a more uniform pattern as a high number of respective genes were generally induced during the whole process of AR formation. The important role of ethylene for stimulating AR formation was demonstrated by the application of inhibitors of ethylene biosynthesis and perception as well as of the precursor aminocyclopropane-1-carboxylic acid, all changing the number and length of AR. A model is proposed showing the putative role of polar auxin transport and resulting auxin accumulation in initiation of subsequent changes in auxin homeostasis and signal perception with a particular role of Aux/IAA expression. These changes might in turn guide the entrance into the different phases of AR formation. Ethylene biosynthesis, which is stimulated by wounding and does probably also respond to other stresses and auxin, acts as important stimulator of AR formation probably via the expression of ethylene responsive transcription factor genes, whereas the timing of different phases seems to be controlled by auxin. PMID

  11. Evaluation of cells and biological reagents for adventitious agents using degenerate primer PCR and massively parallel sequencing.

    PubMed

    McClenahan, Shasta D; Uhlenhaut, Christine; Krause, Philip R

    2014-12-12

    We employed a massively parallel sequencing (MPS)-based approach to test reagents and model cell substrates including Chinese hamster ovary (CHO), Madin-Darby canine kidney (MDCK), African green monkey kidney (Vero), and High Five insect cell lines for adventitious agents. RNA and DNA were extracted either directly from the samples or from viral capsid-enriched preparations, and then subjected to MPS-based non-specific virus detection with degenerate oligonucleotide primer (DOP) PCR. MPS by 454, Illumina MiSeq, and Illumina HiSeq was compared on independent samples. Virus detection using these methods was reproducibly achieved. Unclassified sequences from CHO cells represented cellular sequences not yet submitted to the databases typically used for sequence identification. The sensitivity of MPS-based virus detection was consistent with theoretically expected limits based on dilution of virus in cellular nucleic acids. Capsid preparation increased the number of viral sequences detected. Potential viral sequences were detected in several samples; in each case, these sequences were either artifactual or (based on additional studies) shown not to be associated with replication-competent viruses. Virus-like sequences were more likely to be identified in BLAST searches using virus-specific databases that did not contain cellular sequences. Detected viral sequences included previously described retrovirus and retrovirus-like sequences in CHO, Vero, MDCK and High Five cells, and nodavirus and endogenous bracovirus sequences in High Five insect cells. Bovine viral diarrhea virus, bovine hokovirus, and porcine circovirus sequences were detected in some reagents. A recently described parvo-like virus present in some nucleic acid extraction resins was also identified in cells and extraction controls from some samples. The present study helps to illustrate the potential for MPS-based strategies in evaluating the presence of viral nucleic acids in various sample types

  12. Aspergillus niger Enhance Bioactive Compounds Biosynthesis As Well As Expression of Functional Genes in Adventitious Roots of Glycyrrhiza uralensis Fisch.

    PubMed

    Li, Jing; Wang, Juan; Li, Jinxin; Liu, Dahui; Li, Hongfa; Gao, Wenyuan; Li, Jianli; Liu, Shujie

    2016-02-01

    In the present study, the culture conditions for the accumulation of Glycyrrhiza uralensis adventitious root metabolites in balloon-type bubble bioreactors (BTBBs) have been optimized. The results of the culture showed that the best culture conditions were a cone angle of 90° bioreactor and 0.4-0.6-0.4-vvm aeration volume. Aspergillus niger can be used as a fungal elicitor to enhance the production of defense compounds in plants. With the addition of a fungal elicitor (derived from Aspergillus niger), the maximum accumulation of total flavonoids (16.12 mg g(-1)) and glycyrrhetinic acid (0.18 mg g(-1)) occurred at a dose of 400 mg L(-1) of Aspergillus niger resulting in a 3.47-fold and 1.8-fold increase over control roots. However, the highest concentration of polysaccharide (106.06 mg g(-1)) was achieved with a mixture of elicitors (Aspergillus niger and salicylic acid) added to the medium, resulting in a 1.09-fold increase over Aspergillus niger treatment alone. Electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed, showing that seven compounds were present after treatment with the elicitors, including uralsaponin B, licorice saponin B2, liquiritin, and (3R)-vestitol, only identified in the mixed elicitor treatment group. It has also been found that elicitors (Aspergillus niger and salicylic acid) significantly upregulated the expression of the cinnamate 4-hydroxylase (C4H), β-amyrin synthase (β-AS), squalene epoxidase (SE) and a cytochrome P450 monooxygenase (CYP72A154) genes, which are involved in the biosynthesis of bioactive compounds, and increased superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activity. PMID:26490378

  13. Evaluation of cells and biological reagents for adventitious agents using degenerate primer PCR and massively parallel sequencing.

    PubMed

    McClenahan, Shasta D; Uhlenhaut, Christine; Krause, Philip R

    2014-12-12

    We employed a massively parallel sequencing (MPS)-based approach to test reagents and model cell substrates including Chinese hamster ovary (CHO), Madin-Darby canine kidney (MDCK), African green monkey kidney (Vero), and High Five insect cell lines for adventitious agents. RNA and DNA were extracted either directly from the samples or from viral capsid-enriched preparations, and then subjected to MPS-based non-specific virus detection with degenerate oligonucleotide primer (DOP) PCR. MPS by 454, Illumina MiSeq, and Illumina HiSeq was compared on independent samples. Virus detection using these methods was reproducibly achieved. Unclassified sequences from CHO cells represented cellular sequences not yet submitted to the databases typically used for sequence identification. The sensitivity of MPS-based virus detection was consistent with theoretically expected limits based on dilution of virus in cellular nucleic acids. Capsid preparation increased the number of viral sequences detected. Potential viral sequences were detected in several samples; in each case, these sequences were either artifactual or (based on additional studies) shown not to be associated with replication-competent viruses. Virus-like sequences were more likely to be identified in BLAST searches using virus-specific databases that did not contain cellular sequences. Detected viral sequences included previously described retrovirus and retrovirus-like sequences in CHO, Vero, MDCK and High Five cells, and nodavirus and endogenous bracovirus sequences in High Five insect cells. Bovine viral diarrhea virus, bovine hokovirus, and porcine circovirus sequences were detected in some reagents. A recently described parvo-like virus present in some nucleic acid extraction resins was also identified in cells and extraction controls from some samples. The present study helps to illustrate the potential for MPS-based strategies in evaluating the presence of viral nucleic acids in various sample types

  14. Effects of ion beam irradiation on adventitious shoot regeneration from in vitro leaf explants of Saintpaulia ionahta

    NASA Astrophysics Data System (ADS)

    Zhou, L. B.; Li, W. J.; Ma, S.; Dong, X. C.; Yu, L. X.; Li, Q.; Zhou, G. M.; Gao, Q. X.

    2006-03-01

    The effects of 960 MeV carbon ion beam and 8 MeV X-ray irradiation on adventitious shoots from in vitro leaf explants of two different Saintpaulia ionahta (Mauve and Indikon) cultivars were studied with regard to tissue increase, shoots differentiation and morphology changes in the shoots. The experimental results showed that the survival fraction of shoot formation for the Mauve and Indikon irradiated with the carbon ion beam at 20 Gy were 0.715 and 0.600, respectively, while those for both the cultivars exposed to the X-ray irradiation at the same dose were 1.000. Relative biological effectiveness (RBE) of Mauve with respect to X-ray was about two. Secondly, the percentage of regenerating explants with malformed shoots in all Mauve regenerating explants irradiated with carbon ion beam at 20 Gy accounted for 49.6%, while that irradiated with the same dose of X-ray irradiation was only 4.7%; as for Saintpaulia ionahta Indikon irradiated with 20 Gy carbon ion beam, the percentage was 43.3%, which was higher than that of X-ray irradiation. Last, many chlorophyll deficient and other varieties of mutants were obtained in this study. Based on the results above, it can be concluded that the effect of mutation induction by carbon ion beam irradiation on the leaf explants of Saintpaulia ionahta is better than that by X-ray irradiation; and the optimal mutagenic dose varies from 20 Gy to 25 Gy for carbon ion beam irradiation.

  15. Augmenting in vitro shoot multiplication by vipul (triacontanol) and adventitious rhizogenesis by rice bran extract in Dendrocalamus strictus.

    PubMed

    Mishra, Y; Rana, P K; Shirin, F; Ansari, S A

    2001-02-01

    Like other bamboo species, Dendrocalamus strictus flowers gregariously after a prolonged intermast period of 48 years and constitutes an ideal material for in vitro clonal propagation. In this study, MS liquid medium containing 0.5, 1.0 and 2.0 mL/L vipul (Godrej Agrovet, Ltd., Sachin, India), a commercial formulation of triacontanol, with or without BA (3.0 mg/L) was tested for in vitro shoot multiplication and 1.0, 2.5 and 5.0 mL/L of 20% (w/v) alcoholic/aqueous rice bran extract (alone or in combination) with NAA (3 mg/L) used for in vitro adventitious rhizogenesis in single node culture derived shoots of Dendrocalamus strictus.. After a multiplication cycle for 4-5 week, vipul (0.5 mL/L) with BA (3.0 mg/L) in the culture medium induced 4.59 fold shoot multiplication rate whereas application of BA and vipul alone had corresponding values of 3.29 and 0.53 fold respectively. Maximum vipul concentration (2 mL/L) with BA (3 mg/L) exhibited shoot multiplication higher than (or equal to) that of BA alone. Maximum in vitro rooting percentage (55.66%) was obtained on half MS medium enriched with alcoholic rice bran extract (2.5 mL/L) and NAA (3 mg/L). This is the first investigation reporting amelioration of in vitro shoot multiplication rate by triacontanol and rooting percentage by rice bran extract in explants from mature bamboo culms. The protocol is economical and rapid for in vitro clonal propagation of Dendrocalamus strictus.

  16. Dark exposure of petunia cuttings strongly improves adventitious root formation and enhances carbohydrate availability during rooting in the light.

    PubMed

    Klopotek, Yvonne; Haensch, Klaus-Thomas; Hause, Bettina; Hajirezaei, Mohammad-Reza; Druege, Uwe

    2010-05-01

    The effect of temporary dark exposure on adventitious root formation (ARF) in Petuniaxhybrida 'Mitchell' cuttings was investigated. Histological and metabolic changes in the cuttings during the dark treatment and subsequent rooting in the light were recorded. Excised cuttings were exposed to the dark for seven days at 10 degrees C followed by a nine-day rooting period in perlite or were rooted immediately for 16 days in a climate chamber at 22/20 degrees C (day/night) and a photosynthetic photon flux density (PPFD) of 100micromolm(-2)s(-1). Dark exposure prior to rooting increased, accelerated and synchronized ARF. The rooting period was reduced from 16 days (non-treated cuttings) to 9 days (treated cuttings). Under optimum conditions, despite the reduced rooting period, dark-exposed cuttings produced a higher number and length of roots than non-treated cuttings. An increase in temperature to 20 degrees C during the dark treatment or extending the cold dark exposure to 14 days caused a similar enhancement of root development compared to non-treated cuttings. Root meristem formation had already started during the dark treatment and was enhanced during the subsequent rooting period. Levels of soluble sugars (glucose, fructose and sucrose) and starch in leaf and basal stem tissues significantly decreased during the seven days of dark exposure. This depletion was, however, compensated during rooting after 6 and 24h for soluble sugars in leaves and the basal stem, respectively, whereas the sucrose level in the basal stem was already increased at 6h. The association of higher carbohydrate levels with improved rooting in previously dark-exposed versus non-treated cuttings indicates that increased post-darkness carbohydrate availability and allocation towards the stem base contribute to ARF under the influence of dark treatment and provide energy for cell growth subject to a rising sink intensity in the base of the cutting. PMID:20047776

  17. Promoting Roles of Melatonin in Adventitious Root Development of Solanum lycopersicum L. by Regulating Auxin and Nitric Oxide Signaling

    PubMed Central

    Wen, Dan; Gong, Biao; Sun, Shasha; Liu, Shiqi; Wang, Xiufeng; Wei, Min; Yang, Fengjuan; Li, Yan; Shi, Qinghua

    2016-01-01

    Melatonin (MT) plays integral roles in regulating several biological processes including plant growth, seed germination, flowering, senescence, and stress responses. This study investigated the effects of MT on adventitious root formation (ARF) of de-rooted tomato seedlings. Exogenous MT positively or negatively influenced ARF, which was dependent on the concentration of MT application. In the present experiment, 50 μM MT showed the best effect on inducing ARF. Interestingly, exogenous MT promoted the accumulation of endogenous nitric oxide (NO) by down-regulating the expression of S-nitrosoglutathione reductase (GSNOR). To determine the interaction of MT and NO in ARF, MT synthesis inhibitor p-chlorophenylalanine, NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt as well as GSNOR-overexpression plants with low NO levels were used. The function of MT was removed by NO scavenger or GSNOR-overexpression plants. However, application of MT synthesis inhibitor did little to abolish the function of NO. These results indicate that NO, as a downstream signal, was involved in the MT-induced ARF. Concentrations of indole-3-acetic acid and indole-3-butyric acid, as well as the expression of several genes related to the auxin signaling pathway (PIN1, PIN3, PIN7, IAA19, and IAA24), showed that MT influenced auxin transport and signal transduction as well as auxin accumulation through the NO signaling pathway. Collectively, these strongly suggest that elevated NO levels resulting from inhibited GSNOR activity and auxin signaling were involved in the MT-induced ARF in tomato plants. This can be applied in basic research and breeding. PMID:27252731

  18. Triterpene and Flavonoid Biosynthesis and Metabolic Profiling of Hairy Roots, Adventitious Roots, and Seedling Roots of Astragalus membranaceus.

    PubMed

    Park, Yun Ji; Thwe, Aye Aye; Li, Xiaohua; Kim, Yeon Jeong; Kim, Jae Kwang; Arasu, Mariadhas Valan; Al-Dhabi, Naif Abdullah; Park, Sang Un

    2015-10-14

    Astragalus membranaceus is an important traditional Chinese herb with various medical applications. Astragalosides (ASTs), calycosin, and calycosin-7-O-β-d-glucoside (CG) are the primary metabolic components in A. membranaceus roots. The dried roots of A. membranaceus have various medicinal properties. The present study aimed to investigate the expression levels of genes related to the biosynthetic pathways of ASTs, calycosin, and CG to investigate the differences between seedling roots (SRs), adventitious roots (ARs), and hairy roots (HRs) using quantitative real-time polymerase chain reaction (qRT-PCR). qRT-PCR study revealed that the transcription level of genes involved in the AST biosynthetic pathway was lowest in ARs and showed similar patterns in HRs and SRs. Moreover, most genes involved in the synthesis of calycosin and CG exhibited the highest expression levels in SRs. High-performance liquid chromatography (HPLC) analysis indicated that the expression level of the genes correlated with the content of ASTs, calycosin, and CG in the three different types of roots. ASTs were the most abundant in SRs. CG accumulation was greater than calycosin accumulation in ARs and HRs, whereas the opposite was true in SRs. Additionally, 40 metabolites were identified using gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS). Principal component analysis (PCA) documented the differences among SRs, ARs, and HRs. PCA comparatively differentiated among the three samples. The results of PCA showed that HRs were distinct from ARs and SRs on the basis of the dominant amounts of sugars and clusters derived from closely similar biochemical pathways. Also, ARs had a higher concentration of phenylalanine, a precursor for the phenylpropanoid biosynthetic pathway, as well as CG. TCA cycle intermediates levels including succinic acid and citric acid indicated a higher amount in SRs than in the others.

  19. Comprehensive Transcriptome Analysis Unravels the Existence of Crucial Genes Regulating Primary Metabolism during Adventitious Root Formation in Petunia hybrida

    PubMed Central

    Ahkami, Amirhossein; Scholz, Uwe; Steuernagel, Burkhard; Strickert, Marc; Haensch, Klaus-Thomas; Druege, Uwe; Reinhardt, Didier; Nouri, Eva; von Wirén, Nicolaus; Franken, Philipp; Hajirezaei, Mohammad-Reza

    2014-01-01

    To identify specific genes determining the initiation and formation of adventitious roots (AR), a microarray-based transcriptome analysis in the stem base of the cuttings of Petunia hybrida (line W115) was conducted. A microarray carrying 24,816 unique, non-redundant annotated sequences was hybridized to probes derived from different stages of AR formation. After exclusion of wound-responsive and root-regulated genes, 1,354 of them were identified which were significantly and specifically induced during various phases of AR formation. Based on a recent physiological model distinguishing three metabolic phases in AR formation, the present paper focuses on the response of genes related to particular metabolic pathways. Key genes involved in primary carbohydrate metabolism such as those mediating apoplastic sucrose unloading were induced at the early sink establishment phase of AR formation. Transcriptome changes also pointed to a possible role of trehalose metabolism and SnRK1 (sucrose non-fermenting 1- related protein kinase) in sugar sensing during this early step of AR formation. Symplastic sucrose unloading and nucleotide biosynthesis were the major processes induced during the later recovery and maintenance phases. Moreover, transcripts involved in peroxisomal beta-oxidation were up-regulated during different phases of AR formation. In addition to metabolic pathways, the analysis revealed the activation of cell division at the two later phases and in particular the induction of G1-specific genes in the maintenance phase. Furthermore, results point towards a specific demand for certain mineral nutrients starting in the recovery phase. PMID:24978694

  20. Promoting Roles of Melatonin in Adventitious Root Development of Solanum lycopersicum L. by Regulating Auxin and Nitric Oxide Signaling.

    PubMed

    Wen, Dan; Gong, Biao; Sun, Shasha; Liu, Shiqi; Wang, Xiufeng; Wei, Min; Yang, Fengjuan; Li, Yan; Shi, Qinghua

    2016-01-01

    Melatonin (MT) plays integral roles in regulating several biological processes including plant growth, seed germination, flowering, senescence, and stress responses. This study investigated the effects of MT on adventitious root formation (ARF) of de-rooted tomato seedlings. Exogenous MT positively or negatively influenced ARF, which was dependent on the concentration of MT application. In the present experiment, 50 μM MT showed the best effect on inducing ARF. Interestingly, exogenous MT promoted the accumulation of endogenous nitric oxide (NO) by down-regulating the expression of S-nitrosoglutathione reductase (GSNOR). To determine the interaction of MT and NO in ARF, MT synthesis inhibitor p-chlorophenylalanine, NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt as well as GSNOR-overexpression plants with low NO levels were used. The function of MT was removed by NO scavenger or GSNOR-overexpression plants. However, application of MT synthesis inhibitor did little to abolish the function of NO. These results indicate that NO, as a downstream signal, was involved in the MT-induced ARF. Concentrations of indole-3-acetic acid and indole-3-butyric acid, as well as the expression of several genes related to the auxin signaling pathway (PIN1, PIN3, PIN7, IAA19, and IAA24), showed that MT influenced auxin transport and signal transduction as well as auxin accumulation through the NO signaling pathway. Collectively, these strongly suggest that elevated NO levels resulting from inhibited GSNOR activity and auxin signaling were involved in the MT-induced ARF in tomato plants. This can be applied in basic research and breeding. PMID:27252731

  1. Ginsenoside Production and Morphological Characterization of Wild Ginseng (Panax ginseng Meyer) Mutant Lines Induced by γ-irradiation (60Co) of Adventitious Roots

    PubMed Central

    Zhang, Jun-Ying; Bae, Tae-Woong; Boo, Kyung-Hwan; Sun, Hyeon-Jin; Song, In-Ja; Pham, Chi-Hoa; Ganesan, Markkandan; Yang, Dae-Hwa; Kang, Hong-Gyu; Ko, Suk-Min; Riu, Key-Zung; Lim, Pyung-Ok; Lee, Hyo-Yeon

    2011-01-01

    With the purpose of improving ginsenoside content in adventitious root cultures of Korean wild ginseng (Panax ginseng Meyer), the roots were treated with different dosages of γ-ray (5, 10, 25, 50, 75, 100, and 200 Gy). The growth of adventitious roots was inhibited at over 100 Gy. The irradiated adventitious roots showed significant variation in the morphological parameters and crude saponin content at 50 to100 Gy. Therefore, four mutant cell lines out of the propagation of 35 cell lines treated with 50 Gy and 100 Gy were selected on the basis of phenotypic morphology and crude saponin contents relative to the wild type control. The contents of 7 major ginsenosides (Rg1, Re, Rb1, Rb2, Rc, Rf, and Rd) were determined for cell lines 1 and 3 from 100 Gy and lines 2 and 4 from 50 Gy treatments. Cell line 2 showed more secondary roots, longer length and superior growth rate than the root controls in flasks and bioreactors. Cell line 1 showed larger average diameter and the growth rate in the bioreactor was comparable with that of the control but greater in the flask cultured roots. Cell lines 1 and 2, especially the former, showed much more ginsenoside contents than the control in flasks and bioreactors. Therefore, we chose cell line 1 for further study of ginsenoside contents. The crude saponin content of line 1 in flask and bioreactor cultures increased by 1.4 and 1.8-fold, respectively, compared to the control. Total contents of 7 ginsenoside types (Rg1, Re, Rb1, Rb2, Rc, Rf, and Rd) increased by 1.8 and 2.3-fold, respectively compared to the control. Crude saponin and ginsenoside contents in the bioreactor culture increased by about 1.4-fold compared to that the flask culture. PMID:23717071

  2. Enhancement of Anti-Inflammatory Activity of Aloe vera Adventitious Root Extracts through the Alteration of Primary and Secondary Metabolites via Salicylic Acid Elicitation

    PubMed Central

    Lee, Yun Sun; Ju, Hyun Kyoung; Kim, Yeon Jeong; Lim, Tae-Gyu; Uddin, Md Romij; Kim, Yeon Bok; Baek, Jin Hong; Kwon, Sung Won; Lee, Ki Won; Seo, Hak Soo; Park, Sang Un; Yang, Tae-Jin

    2013-01-01

    Aloe vera (Asphodeloideae) is a medicinal plant in which useful secondary metabolites are plentiful. Among the representative secondary metabolites of Aloe vera are the anthraquinones including aloe emodin and chrysophanol, which are tricyclic aromatic quinones synthesized via a plant-specific type III polyketide biosynthesis pathway. However, it is not yet clear which cellular responses can induce the pathway, leading to production of tricyclic aromatic quinones. In this study, we examined the effect of endogenous elicitors on the type III polyketide biosynthesis pathway and identified the metabolic changes induced in elicitor-treated Aloe vera adventitious roots. Salicylic acid, methyl jasmonate, and ethephon were used to treat Aloe vera adventitious roots cultured on MS liquid media with 0.3 mg/L IBA for 35 days. Aloe emodin and chrysophanol were remarkably increased by the SA treatment, more than 10–11 and 5–13 fold as compared with untreated control, respectively. Ultra-performance liquid chromatography-electrospray ionization mass spectrometry analysis identified a total of 37 SA-induced compounds, including aloe emodin and chrysophanol, and 3 of the compounds were tentatively identified as tricyclic aromatic quinones. Transcript accumulation analysis of polyketide synthase genes and gas chromatography mass spectrometry showed that these secondary metabolic changes resulted from increased expression of octaketide synthase genes and decreases in malonyl-CoA, which is the precursor for the tricyclic aromatic quinone biosynthesis pathway. In addition, anti-inflammatory activity was enhanced in extracts of SA-treated adventitious roots. Our results suggest that SA has an important role in activation of the plant specific-type III polyketide biosynthetic pathway, and therefore that the efficacy of Aloe vera as medicinal agent can be improved through SA treatment. PMID:24358188

  3. Enhancement of anti-inflammatory activity of Aloe vera adventitious root extracts through the alteration of primary and secondary metabolites via salicylic acid elicitation.

    PubMed

    Lee, Yun Sun; Ju, Hyun Kyoung; Kim, Yeon Jeong; Lim, Tae-Gyu; Uddin, Md Romij; Kim, Yeon Bok; Baek, Jin Hong; Kwon, Sung Won; Lee, Ki Won; Seo, Hak Soo; Park, Sang Un; Yang, Tae-Jin

    2013-01-01

    Aloe vera (Asphodeloideae) is a medicinal plant in which useful secondary metabolites are plentiful. Among the representative secondary metabolites of Aloe vera are the anthraquinones including aloe emodin and chrysophanol, which are tricyclic aromatic quinones synthesized via a plant-specific type III polyketide biosynthesis pathway. However, it is not yet clear which cellular responses can induce the pathway, leading to production of tricyclic aromatic quinones. In this study, we examined the effect of endogenous elicitors on the type III polyketide biosynthesis pathway and identified the metabolic changes induced in elicitor-treated Aloe vera adventitious roots. Salicylic acid, methyl jasmonate, and ethephon were used to treat Aloe vera adventitious roots cultured on MS liquid media with 0.3 mg/L IBA for 35 days. Aloe emodin and chrysophanol were remarkably increased by the SA treatment, more than 10-11 and 5-13 fold as compared with untreated control, respectively. Ultra-performance liquid chromatography-electrospray ionization mass spectrometry analysis identified a total of 37 SA-induced compounds, including aloe emodin and chrysophanol, and 3 of the compounds were tentatively identified as tricyclic aromatic quinones. Transcript accumulation analysis of polyketide synthase genes and gas chromatography mass spectrometry showed that these secondary metabolic changes resulted from increased expression of octaketide synthase genes and decreases in malonyl-CoA, which is the precursor for the tricyclic aromatic quinone biosynthesis pathway. In addition, anti-inflammatory activity was enhanced in extracts of SA-treated adventitious roots. Our results suggest that SA has an important role in activation of the plant specific-type III polyketide biosynthetic pathway, and therefore that the efficacy of Aloe vera as medicinal agent can be improved through SA treatment.

  4. Thalidomide influences atherogenesis in aortas of ApoE(-/-)/LDLR (-/-) double knockout mice: a nano-CT study.

    PubMed

    Kampschulte, Marian; Gunkel, Irina; Stieger, Philipp; Sedding, Daniel G; Brinkmann, Anne; Ritman, Erik L; Krombach, Gabriele A; Langheinrich, Alexander C

    2014-04-01

    Plaque progression in atherosclerosis is closely connected to angiogenesis due to vasa vasorum (VV) growth. Objective of this study was to determine the unknown long-term effect of thalidomide on adventitial VV neovascularization and plaque progression using nano-focussed computed tomography (nano-CT). Proliferation and migration assays in human coronary artery endothelial cells (HCAEC) measured number of viable cells after incubation with thalidomide. Male ApoE(-/-)/LDLR(-/-) (AL) mice (n = 5) received a thalidomide containing western diet (WD) over 29 weeks. Another five male AL mice (WD without thalidomide) served as control group. Descending aortas were scanned with nano-CT at (1.5 μm)(3) isotropic voxel size. Number and area of adventitial VV as well as plaque cross sectional area were measured. Results were complemented by histology. Thalidomide inhibited proliferation and migration of HCAEC dose-dependently. VV neovascularization decreased in number per cross section (7.66 ± 0.301 vs. 8.62 ± 0.164, p < 0.001) and in cross sectional area (0.0183 ± 0.0011 vs. 0.0238 ± 0.0008 mm(2), p < 0.001). Cross sectional area of plaque decreased significantly when treated with thalidomide (0.57 ± 0.0187 vs. 0.803 ± 0.0148 mm(2), p < 0.001). Nano-CT imaging revealed a reduced plaque growth and VV neovascularization after long-term application of thalidomide. Therefore, nano-CT can be considered as a new method to detect therapeutic effects in experimental models of atherosclerosis.

  5. Cystic adventitial disease of the popliteal artery: elongation into the media of the popliteal artery and communication with the knee joint capsule: report of a case.

    PubMed

    Unno, N; Kaneko, H; Uchiyama, T; Yamamoto, N; Nakamura, S

    2000-01-01

    Cystic disease of the popliteal artery is a rare disorder in which most cases involve the formation of an adventitial cyst that disturbs the popliteal artery blood flow. We present herein the case of a patient presenting with popliteal artery occlusion due to compression by a cyst which formed at the media of the popliteal artery. The onset occurred during a baseball game in which he played catcher. Preoperative magnetic resonance imaging demonstrated a communication of the cyst with the adjacent knee joint. This unusual case could provide important clues to help identify the pathogenesis of this disease.

  6. Adventitial Alterations Are the Main Features in Pulmonary Artery Remodeling due to Long-Term Chronic Intermittent Hypobaric Hypoxia in Rats

    PubMed Central

    Brito, Julio; Siques, Patricia; Arribas, Silvia M.; López de Pablo, Angel L.; González, M. Carmen; Naveas, Nelson; Flores, Karen; León-Velarde, Fabiola; Pulido, Ruth; Ordenes, Stefany; López, M. Rosario

    2015-01-01

    Long-term chronic intermittent exposure to altitude hypoxia is a labor phenomenon requiring further research. Using a rat model, we examined whether this type of exposure differed from chronic exposure in terms of pulmonary artery remodeling and other features. Rats were subjected to chronic hypoxia (CH, n = 9) and long-term intermittent hypoxia (CIH2x2; 2 days of hypoxia/2 days of normoxia, n = 10) in a chamber (428 Torr, 4,600 m of altitude) for 46 days and compared to rats under normoxia (NX, n = 10). Body weight, hematocrit, and right ventricle ratio were measured. Pulmonary artery remodeling was assessed using confocal microscopy of tissues stained with a nuclear dye (DAPI) and CD11b antibody. Both hypoxic conditions exhibited increased hematocrit and hypertrophy of the right ventricle, tunica adventitia, and tunica media, with no changes in lumen size. The medial hypertrophy area (larger in CH) depicted a significant increase in smooth muscle cell number. Additionally, CIH2x2 increased the adventitial hypertrophy area, with an increased cellularity and a larger prevalence of clustered inflammatory cells. In conclusion, CIH2x2 elicits milder effects on pulmonary artery medial layer muscularization and subsequent right ventricular hypertrophy than CH. However, CIH2x2 induces greater and characteristic alterations of the adventitial layer. PMID:25738150

  7. Adventitial alterations are the main features in pulmonary artery remodeling due to long-term chronic intermittent hypobaric hypoxia in rats.

    PubMed

    Brito, Julio; Siques, Patricia; Arribas, Silvia M; López de Pablo, Angel L; González, M Carmen; Naveas, Nelson; Arriaza, Karem; Flores, Karen; León-Velarde, Fabiola; Pulido, Ruth; Ordenes, Stefany; López, M Rosario

    2015-01-01

    Long-term chronic intermittent exposure to altitude hypoxia is a labor phenomenon requiring further research. Using a rat model, we examined whether this type of exposure differed from chronic exposure in terms of pulmonary artery remodeling and other features. Rats were subjected to chronic hypoxia (CH, n = 9) and long-term intermittent hypoxia (CIH2x2; 2 days of hypoxia/2 days of normoxia, n = 10) in a chamber (428 Torr, 4,600 m of altitude) for 46 days and compared to rats under normoxia (NX, n = 10). Body weight, hematocrit, and right ventricle ratio were measured. Pulmonary artery remodeling was assessed using confocal microscopy of tissues stained with a nuclear dye (DAPI) and CD11b antibody. Both hypoxic conditions exhibited increased hematocrit and hypertrophy of the right ventricle, tunica adventitia, and tunica media, with no changes in lumen size. The medial hypertrophy area (larger in CH) depicted a significant increase in smooth muscle cell number. Additionally, CIH2x2 increased the adventitial hypertrophy area, with an increased cellularity and a larger prevalence of clustered inflammatory cells. In conclusion, CIH2x2 elicits milder effects on pulmonary artery medial layer muscularization and subsequent right ventricular hypertrophy than CH. However, CIH2x2 induces greater and characteristic alterations of the adventitial layer.

  8. Direct reprogramming of adult somatic cells toward adventitious root formation in forest tree species: the effect of the juvenile–adult transition

    PubMed Central

    Díaz-Sala, Carmen

    2014-01-01

    Cellular plasticity refers, among others, to the capability of differentiated cells to switch the differentiation process and acquire new fates. One way by which plant cell plasticity is manifested is through de novo regeneration of organs from somatic differentiated cells in an ectopic location. However, switching the developmental program of adult cells prior to organ regeneration is difficult in many plant species, especially in forest tree species. In these species, a decline in the capacity to regenerate shoots, roots, or embryos from somatic differentiated cells is associated with tree age and maturation. The decline in the ability to form adventitious roots from stem cuttings is one of the most dramatic effects of maturation, and has been the subject of investigations on the basic nature of the process. Cell fate switches, both in plants and animals, are characterized by remarkable changes in the pattern of gene expression, as cells switch from the characteristic expression pattern of a somatic cell to a new one directing a new developmental pathway. Therefore, determining the way by which cells reset their gene expression pattern is crucial to understand cellular plasticity. The presence of specific cellular signaling pathways or tissue-specific factors underlying the establishment, maintenance, and redirection of gene expression patterns in the tissues involved in adventitious root formation could be crucial for cell fate switch and for the control of age-dependent cellular plasticity. PMID:25071793

  9. In Vitro Culture Conditions and OeARF and OeH3 Expressions Modulate Adventitious Root Formation from Oleaster (Olea europaea L. subsp. europaea var. sylvestris) Cuttings

    PubMed Central

    Gagliardi, Cinzia; Bruno, Leonardo; Bitonti, Maria Beatrice

    2014-01-01

    Olea europaea L. subsp. europaea var. sylvestris, also named oleaster, is the wild form of olive and it is used as rootstock and pollen donor for many cultivated varieties. An efficient procedure for in vitro propagation of oleaster was established in this study. A zeatin concentration of 2.5 mg/L was effective to induce an appreciable vegetative growth. Also high rooting efficiency was obtained by using a short IBA pulse, followed by two different IBA concentrations in the culture medium. With the aim to enlarge knowledge on the molecular aspects of adventitious rooting, we also evaluated the transcriptional modulation of an ARFs member and HISTONE H3 genes, involved in auxin signaling and cell replication, respectively, during the root induction phase of cuttings. The obtained results suggest that the selected genes, as markers of the induction phase, could be very useful for setting up efficient culture conditions along the rooting process, thus increasing micropropagation efficiency. PMID:24587768

  10. In vitro culture conditions and OeARF and OeH3 expressions modulate adventitious root formation from oleaster (Olea europaea L. subsp. europaea var. sylvestris) cuttings.

    PubMed

    Chiappetta, Adriana; Gagliardi, Cinzia; Bruno, Leonardo; Bitonti, Maria Beatrice

    2014-01-01

    Olea europaea L. subsp. europaea var. sylvestris, also named oleaster, is the wild form of olive and it is used as rootstock and pollen donor for many cultivated varieties. An efficient procedure for in vitro propagation of oleaster was established in this study. A zeatin concentration of 2.5 mg/L was effective to induce an appreciable vegetative growth. Also high rooting efficiency was obtained by using a short IBA pulse, followed by two different IBA concentrations in the culture medium. With the aim to enlarge knowledge on the molecular aspects of adventitious rooting, we also evaluated the transcriptional modulation of an ARFs member and HISTONE H3 genes, involved in auxin signaling and cell replication, respectively, during the root induction phase of cuttings. The obtained results suggest that the selected genes, as markers of the induction phase, could be very useful for setting up efficient culture conditions along the rooting process, thus increasing micropropagation efficiency. PMID:24587768

  11. Systematic Evaluation of In Vitro and In Vivo Adventitious Virus Assays for the Detection of Viral Contamination of Cell Banks and Biological Products1

    PubMed Central

    Gombold, James; Karakasidis, Stephen; Niksa, Paula; Podczasy, John; Neumann, Kitti; Richardson, James; Sane, Nandini; Johnson-Leva, Renita; Randolph, Valerie; Sadoff, Jerald; Minor, Phillip; Schmidt, Alexander; Duncan, Paul; Sheets, Rebecca L.

    2015-01-01

    Viral vaccines and the cell substrates used to manufacture them are subjected to tests for adventitious agents, including viruses, which might contaminant them. Some of the compendial methods (in vivo and in vitro in cell culture) were established in the mid-20th century. These methods have not been subjected to current assay validation, as new methods would need to be. This study was undertaken to provide insight into the breadth (selectivity) and sensitivity (limit of detection) of the routine methods, two such validation parameters. Sixteen viral stocks were prepared and characterized. These stocks were tested in serial dilutions by the routine methods to establish which viruses were detected by which methods and above what limit of detection. Sixteen out of sixteen viruses were detected in vitro, though one (bovine viral diarrhea virus) required special conditions to detect and another (rubella virus) was detected with low sensitivity. Many were detected at levels below 1 TCID50 or PFU (titers were established on the production cell line in most cases). In contrast, in vivo, only 6/11 viruses were detected, and 4 of these were detected only at amounts one or more logs above 1 TCID50 or PFU. Only influenza virus and vesicular stomatitis virus were detected at lower amounts in vivo than in vitro. Given the call to reduce, refine, or replace (3 R's) the use of animals in product safety testing and the emergence of new technologies for the detection of viruses, a re-examination of the current adventitious virus testing strategies seems warranted. Suggested pathways forward are offered. PMID:24681273

  12. Methods to select suitable fetal bovine serum for use in quality control assays for the detection of adventitious viruses from biological products.

    PubMed

    Kozasa, Takashi; Aoki, Hiroshi; Nakajima, Nao; Fukusho, Akio; Ishimaru, Masatoshi; Nakamura, Shigeyuki

    2011-07-01

    Production of biological products, especially vaccines, usually requires materials derived from animals, and there are always risks that animal pathogens derived from these materials could contaminate the final products. Detection of adventitious agents is performed by quality control tests. In these biological assays, animal derived materials are also used and another problem arises, as fetal bovine serum (FBS) is used as an ingredient in tissue culture media. FBS contaminated with bovine viral diarrhea virus (BVDV) or other bovine pathogens, as well as antibodies against these pathogens may lead to false results in quality control assays. In this study, in order to determine the actual status of commercial FBS, we performed quality tests on various FBS samples. As a result, in 28 of 49 FBS samples (57.1%), pestivirus genes were detected by pan-pestivirus reverse transcription-polymerase chain reaction assay. Furthermore, two samples contained infectious BVDV. Neutralizing antibodies against BVDVs were detected in 48 of 49 samples (97.6%) by the virus neutralization test based on the serum-dilution or virus-dilution methods. Antibodies against other bovine pathogens were detected rarely in these samples. From our results, we recommend methods to select FBS that are focused on detection of BVDV and neutralizing antibodies against BVDV.

  13. Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by γ-irradiation ((60)Co) of adventitious roots.

    PubMed

    Zhang, Jun-Ying; Sun, Hyeon-Jin; Song, In-Ja; Bae, Tae-Woong; Kang, Hong-Gyu; Ko, Suk-Min; Kwon, Yong-Ik; Kim, Il-Woung; Lee, Jaechun; Park, Shin-Young; Lim, Pyung-Ok; Kim, Yong Hwan; Lee, Hyo-Yeon

    2014-07-01

    An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

  14. Transgenic manipulation of plant embryo sacs tracked through cell-type-specific fluorescent markers: cell labeling, cell ablation, and adventitious embryos.

    PubMed

    Lawit, Shai J; Chamberlin, Mark A; Agee, April; Caswell, Eric S; Albertsen, Marc C

    2013-06-01

    Expression datasets relating to the Arabidopsis female gametophyte have enabled the creation of a tool set which allows simultaneous visual tracking of each specific cell type (egg, synergids, central cell, and antipodals). This cell-specific, fluorescent labeling tool-set functions from gametophyte cellularization through fertilization and early embryo development. Using this system, cell fates were tracked within Arabidopsis ovules following molecular manipulations, such as the ablation of the egg and/or synergids. Upon egg cell ablation, it was observed that a synergid can switch its developmental fate to become egg/embryo-like upon loss of the native egg. Also, manipulated was the fate of the somatic ovular cells, which can become egg- and embryo-like, reminiscent of adventitious embryony. These advances represent initial steps toward engineering synthetic apomixis resulting in seed derived wholly from the maternal plant. The end goal of applied apomixis research, fixing important agronomic traits such as hybrid vigor, would be a key benefit to agricultural productivity.

  15. A higher sink competitiveness of the rooting zone and invertases are involved in dark stimulation of adventitious root formation in Petunia hybrida cuttings.

    PubMed

    Klopotek, Yvonne; Franken, Philipp; Klaering, Hans-Peter; Fischer, Kerstin; Hause, Bettina; Hajirezaei, Mohammad-Reza; Druege, Uwe

    2016-02-01

    The contribution of carbon assimilation and allocation and of invertases to the stimulation of adventitious root formation in response to a dark pre-exposure of petunia cuttings was investigated, considering the rooting zone (stem base) and the shoot apex as competing sinks. Dark exposure had no effect on photosynthesis and dark respiration during the subsequent light period, but promoted dry matter partitioning to the roots. Under darkness, higher activities of cytosolic and vacuolar invertases were maintained in both tissues when compared to cuttings under light. This was partially associated with higher RNA levels of respective genes. However, activity of cell wall invertases and transcript levels of one cell wall invertase isogene increased specifically in the stem base during the first two days after cutting excision under both light and darkness. During five days after excision, RNA accumulation of four invertase genes indicated preferential expression in the stem base compared to the apex. Darkness shifted the balance of expression of one cytosolic and two vacuolar invertase genes towards the stem base. The results indicate that dark exposure before planting enhances the carbon sink competitiveness of the rooting zone and that expression and activity of invertases contribute to the shift in carbon allocation. PMID:26795147

  16. Comparative Proteomic Analysis Provides Insight into the Key Proteins Involved in Cucumber (Cucumis sativus L.) Adventitious Root Emergence under Waterlogging Stress

    PubMed Central

    Xu, Xuewen; Ji, Jing; Ma, Xiaotian; Xu, Qiang; Qi, Xiaohua; Chen, Xuehao

    2016-01-01

    Waterlogging is a common abiotic stress in both natural and agricultural systems, and it primarily affects plant growth by the slow oxygen diffusion in water. To sustain root function in the hypoxic environment, a key adaptation for waterlogging tolerant plants is the formation of adventitious roots (ARs). We found that cucumber waterlogging tolerant line Zaoer-N seedlings adapt to waterlogging stress by developing a larger number of ARs in hypocotyls, while almost no AR is generated in sensitive line Pepino. To understand the molecular mechanisms underlying AR emergence, the iTRAQ-based quantitative proteomics approach was employed to map the proteomes of hypocotyls cells of the Zaoer-N and Pepino under control and waterlogging conditions. A total of 5508 proteins were identified and 146 were differentially regulated proteins (DRPs), of which 47 and 56 DRPs were specific to tolerant and sensitive line, respectively. In the waterlogged Zaoer-N hypocotyls, DRPs related to alcohol dehydrogenases (ADH), 1-aminocyclopropane-1-carboxylicacid oxidases, peroxidases, 60S ribosomal proteins, GSDL esterases/lipases, histone deacetylases, and histone H5 and were strongly overrepresented to manage the energy crisis, promote ethylene release, minimize oxidative damage, mobilize storage lipids, and stimulate cell division, differentiation and growth. The evaluations of ethylene production, ADH activity, pyruvate decarboxylase (PDC) activity and ethanol production were in good agreement with the proteomic results. qRT-PCR analysis of the corresponding 146 genes further confirmed the accuracy of the observed protein abundance. These findings shed light on the mechanisms underlying waterlogging triggered cucumber ARs emergence, and provided valuable information for the breeding of cucumber with enhanced tolerance to waterlogging. PMID:27790230

  17. Analysis of Microtubule-Associated-Proteins during IBA-Mediated Adventitious Root Induction Reveals KATANIN Dependent and Independent Alterations of Expression Patterns

    PubMed Central

    Abu-Abied, Mohamad; Mordehaev, Inna; Sunil Kumar, Gujulla B; Ophir, Ron; Wasteneys, Geoffrey O.; Sadot, Einat

    2015-01-01

    Adventitious roots (AR) are post embryonic lateral organs that differentiate from non-root tissues. The understanding of the molecular mechanism which underlies their differentiation is important because of their central role in vegetative plant propagation. Here it was studied how the expression of different microtubule (MT)-associated proteins (MAPs) is affected during AR induction, and whether expression differences are dependent on MT organization itself. To examine AR formation when MTs are disturbed we used two mutants in the MT severing protein KATANIN. It was found that rate and number of AR primordium formed following IBA induction for three days was reduced in bot1-1 and bot1-7 plants. The reduced capacity to form ARs in bot1-1 was associated with altered expression of MAP-encoding genes along AR induction. While the expression of MAP65-4, MAP65-3, AURORA1, AURORA2 and TANGLED, increased in wild-type but not in bot1-1 plants, the expression of MAP65-8 and MDP25 decreased in wild type plants but not in the bot1-1 plant after two days of IBA-treatment. The expression of MOR1 was increased two days after AR induction in wild type and bot1-1 plants. To examine its expression specifically in AR primordium, MOR1 upstream regulatory sequence was isolated and cloned to regulate GFP. Expression of GFP was induced in the primary root tips and lateral roots, in the pericycle of the hypocotyls and in all stages of AR primordium formation. It is concluded that the expression of MAPs is regulated along AR induction and that reduction in KATANIN expression inhibits AR formation and indirectly influences the specific expression of some MAPs. PMID:26630265

  18. Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia.

    PubMed

    Carrabba, M; De Maria, C; Oikawa, A; Reni, C; Rodriguez-Arabaolaza, I; Spencer, H; Slater, S; Avolio, E; Dang, Z; Spinetti, G; Madeddu, P; Vozzi, G

    2016-03-01

    Cell therapy represents a promising option for revascularization of ischemic tissues. However, injection of dispersed cells is not optimal to ensure precise homing into the recipient's vasculature. Implantation of cell-engineered scaffolds around the occluded artery may obviate these limitations. Here, we employed the synthetic polymer polycaprolactone for fabrication of 3D woodpile- or channel-shaped scaffolds by a computer-assisted writing system (pressure assisted micro-syringe square), followed by deposition of gelatin (GL) nanofibers by electro-spinning. Scaffolds were then cross-linked with natural (genipin, GP) or synthetic (3-glycidyloxy-propyl-trimethoxy-silane, GPTMS) agents to improve mechanical properties and durability in vivo. The composite scaffolds were next fixed by crown inserts in each well of a multi-well plate and seeded with adventitial progenitor cells (APCs, 3 cell lines in duplicate), which were isolated/expanded from human saphenous vein surgical leftovers. Cell density, alignment, proliferation and viability were assessed 1 week later. Data from in vitro assays showed channel-shaped/GPTMS-crosslinked scaffolds confer APCs with best alignment and survival/growth characteristics. Based on these results, channel-shaped/GPTMS-crosslinked scaffolds with or without APCs were implanted around the femoral artery of mice with unilateral limb ischemia. Perivascular implantation of scaffolds accelerated limb blood flow recovery, as assessed by laser Doppler or fluorescent microspheres, and increased arterial collaterals around the femoral artery and in limb muscles compared with non-implanted controls. Blood flow recovery and perivascular arteriogenesis were additionally incremented by APC-engineered scaffolds. In conclusion, perivascular application of human APC-engineered scaffolds may represent a novel option for targeted delivery of therapeutic cells in patients with critical limb ischemia. PMID:27011300

  19. Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia.

    PubMed

    Carrabba, M; De Maria, C; Oikawa, A; Reni, C; Rodriguez-Arabaolaza, I; Spencer, H; Slater, S; Avolio, E; Dang, Z; Spinetti, G; Madeddu, P; Vozzi, G

    2016-03-01

    Cell therapy represents a promising option for revascularization of ischemic tissues. However, injection of dispersed cells is not optimal to ensure precise homing into the recipient's vasculature. Implantation of cell-engineered scaffolds around the occluded artery may obviate these limitations. Here, we employed the synthetic polymer polycaprolactone for fabrication of 3D woodpile- or channel-shaped scaffolds by a computer-assisted writing system (pressure assisted micro-syringe square), followed by deposition of gelatin (GL) nanofibers by electro-spinning. Scaffolds were then cross-linked with natural (genipin, GP) or synthetic (3-glycidyloxy-propyl-trimethoxy-silane, GPTMS) agents to improve mechanical properties and durability in vivo. The composite scaffolds were next fixed by crown inserts in each well of a multi-well plate and seeded with adventitial progenitor cells (APCs, 3 cell lines in duplicate), which were isolated/expanded from human saphenous vein surgical leftovers. Cell density, alignment, proliferation and viability were assessed 1 week later. Data from in vitro assays showed channel-shaped/GPTMS-crosslinked scaffolds confer APCs with best alignment and survival/growth characteristics. Based on these results, channel-shaped/GPTMS-crosslinked scaffolds with or without APCs were implanted around the femoral artery of mice with unilateral limb ischemia. Perivascular implantation of scaffolds accelerated limb blood flow recovery, as assessed by laser Doppler or fluorescent microspheres, and increased arterial collaterals around the femoral artery and in limb muscles compared with non-implanted controls. Blood flow recovery and perivascular arteriogenesis were additionally incremented by APC-engineered scaffolds. In conclusion, perivascular application of human APC-engineered scaffolds may represent a novel option for targeted delivery of therapeutic cells in patients with critical limb ischemia.

  20. Concerted transcription of auxin and carbohydrate homeostasis-related genes underlies improved adventitious rooting of microcuttings derived from far-red treated Eucalyptus globulus Labill mother plants.

    PubMed

    Ruedell, Carolina Michels; de Almeida, Márcia Rodrigues; Fett-Neto, Arthur Germano

    2015-12-01

    Economically important plant species, such as Eucalyptus globulus, are often rooting recalcitrant. We have previously shown that far-red light enrichment applied to E. globulus donor-plants improved microcutting rooting competence and increased rooting zone/shoot carbohydrate ratio. To better understand this developmental response, the relative expression profiles of genes involved in auxin signaling (ARF6, ARF8, AGO1), biosynthesis (YUC3) and transport (AUX1, PIN1, PIN2); sucrose cleavage (SUS1, CWINV1), transport (SUC5), hexose phosphorylation (HXK1, FLN1) and starch biosynthesis (SS3) were quantified during adventitious rooting of E. globulus microcuttings derived from donor plants exposed to far-red or white light. Expression of auxin transport-related genes increased in the first days of root induction. Far-red enrichment of donor plants induced ARF6, ARF8 and AGO1 in microcuttings. The first two gene products could activate GH3 and other rooting related genes, whereas AGO1 deregulation of the repressor ARF17 may relief rooting inhibition. Increased sink strength at the basal stem with sucrose unloading in root tissue mediated by SUC and subsequent hydrolysis by SUS1 were also supported by gene expression profile. Fructose phosphorylation and starch biosynthesis could also contribute to proper carbon allocation at the site of rooting, as evidenced by increased expression of related genes. These data are in good agreement with increased contents of hexoses and starch at the cutting base severed from far-red exposed donor plants. To sum up, pathways integrating auxin and carbohydrate metabolism were activated in microcuttings derived from donor plants exposed to far red light enrichment, thereby improving rooting response in E. globulus.

  1. The Tomato (Solanum Lycopersicum cv. Micro-Tom) Natural Genetic Variation Rg1 and the DELLA Mutant Procera Control the Competence Necessary to Form Adventitious Roots and Shoots

    PubMed Central

    Peres, Lázaro Eustáquio Pereira

    2012-01-01

    Despite the wide use of plant regeneration for biotechnological purposes, the signals that allow cells to become competent to assume different fates remain largely unknown. Here, it is demonstrated that the Regeneration1 (Rg1) allele, a natural genetic variation from the tomato wild relative Solanum peruvianum, increases the capacity to form both roots and shoots in vitro; and that the gibberellin constitutive mutant procera (pro) presented the opposite phenotype, reducing organogenesis on either root-inducing medium (RIM) or shoot-inducing medium (SIM). Mutants showing alterations in the formation of specific organs in vitro were the auxin low-sensitivity diageotropica (dgt), the lateral suppresser (ls), and the KNOX-overexpressing Mouse ears (Me). dgt failed to form roots on RIM, Me increased shoot formation on SIM, and the high capacity for in vitro shoot formation of ls contrasted with its recalcitrance to form axillary meristems. Interestingly, Rg1 rescued the in vitro organ formation capacity in proRg1 and dgtRg1 double mutants and the ex vitro low lateral shoot formation in pro and ls. Such epistatic interactions were also confirmed in gene expression and histological analyses conducted in the single and double mutants. Although Me phenocopied the high shoot formation of Rg1 on SIM, it failed to increase rooting on RIM and to rescue the non-branching phenotype of ls. Taken together, these results suggest REGENERATION1 and the DELLA mutant PROCERA as controlling a common competence to assume distinct cell fates, rather than the specific induction of adventitious roots or shoots, which is controlled by DIAGEOTROPICA and MOUSE EARS, respectively. PMID:22915742

  2. Analysis of Microtubule-Associated-Proteins during IBA-Mediated Adventitious Root Induction Reveals KATANIN Dependent and Independent Alterations of Expression Patterns.

    PubMed

    Abu-Abied, Mohamad; Mordehaev, Inna; Sunil Kumar, Gujulla B; Ophir, Ron; Wasteneys, Geoffrey O; Sadot, Einat

    2015-01-01

    Adventitious roots (AR) are post embryonic lateral organs that differentiate from non-root tissues. The understanding of the molecular mechanism which underlies their differentiation is important because of their central role in vegetative plant propagation. Here it was studied how the expression of different microtubule (MT)-associated proteins (MAPs) is affected during AR induction, and whether expression differences are dependent on MT organization itself. To examine AR formation when MTs are disturbed we used two mutants in the MT severing protein KATANIN. It was found that rate and number of AR primordium formed following IBA induction for three days was reduced in bot1-1 and bot1-7 plants. The reduced capacity to form ARs in bot1-1 was associated with altered expression of MAP-encoding genes along AR induction. While the expression of MAP65-4, MAP65-3, AURORA1, AURORA2 and TANGLED, increased in wild-type but not in bot1-1 plants, the expression of MAP65-8 and MDP25 decreased in wild type plants but not in the bot1-1 plant after two days of IBA-treatment. The expression of MOR1 was increased two days after AR induction in wild type and bot1-1 plants. To examine its expression specifically in AR primordium, MOR1 upstream regulatory sequence was isolated and cloned to regulate GFP. Expression of GFP was induced in the primary root tips and lateral roots, in the pericycle of the hypocotyls and in all stages of AR primordium formation. It is concluded that the expression of MAPs is regulated along AR induction and that reduction in KATANIN expression inhibits AR formation and indirectly influences the specific expression of some MAPs. PMID:26630265

  3. Concerted transcription of auxin and carbohydrate homeostasis-related genes underlies improved adventitious rooting of microcuttings derived from far-red treated Eucalyptus globulus Labill mother plants.

    PubMed

    Ruedell, Carolina Michels; de Almeida, Márcia Rodrigues; Fett-Neto, Arthur Germano

    2015-12-01

    Economically important plant species, such as Eucalyptus globulus, are often rooting recalcitrant. We have previously shown that far-red light enrichment applied to E. globulus donor-plants improved microcutting rooting competence and increased rooting zone/shoot carbohydrate ratio. To better understand this developmental response, the relative expression profiles of genes involved in auxin signaling (ARF6, ARF8, AGO1), biosynthesis (YUC3) and transport (AUX1, PIN1, PIN2); sucrose cleavage (SUS1, CWINV1), transport (SUC5), hexose phosphorylation (HXK1, FLN1) and starch biosynthesis (SS3) were quantified during adventitious rooting of E. globulus microcuttings derived from donor plants exposed to far-red or white light. Expression of auxin transport-related genes increased in the first days of root induction. Far-red enrichment of donor plants induced ARF6, ARF8 and AGO1 in microcuttings. The first two gene products could activate GH3 and other rooting related genes, whereas AGO1 deregulation of the repressor ARF17 may relief rooting inhibition. Increased sink strength at the basal stem with sucrose unloading in root tissue mediated by SUC and subsequent hydrolysis by SUS1 were also supported by gene expression profile. Fructose phosphorylation and starch biosynthesis could also contribute to proper carbon allocation at the site of rooting, as evidenced by increased expression of related genes. These data are in good agreement with increased contents of hexoses and starch at the cutting base severed from far-red exposed donor plants. To sum up, pathways integrating auxin and carbohydrate metabolism were activated in microcuttings derived from donor plants exposed to far red light enrichment, thereby improving rooting response in E. globulus. PMID:26397200

  4. Tuberculous iliac artery aneurysm in a pediatric patient.

    PubMed

    Cox, Sharon Gail; Naidoo, Nadraj G; Wood, Richard John; Clark, Lizelle; Kilborn, Tracy

    2013-03-01

    Vascular complications of tuberculous infections are rare and occur even less frequently in the pediatric population. Tuberculous pseudoaneurysms can occur either as a result of contiguous spread from a neighboring focus-invariably infected lymph nodes-or by hematogenous spread and seeding of acid-fast bacilli that lodge in the adventitia or media via the vasa vasorum. We report a case of turberculous right common iliac artery pseudoaneurysm in a 12-year-old and review the relevant literature.

  5. Molecular cloning and characterization of the genes encoding an auxin efflux carrier and the auxin influx carriers associated with the adventitious root formation in mango (Mangifera indica L.) cotyledon segments.

    PubMed

    Li, Yun-He; Zou, Ming-Hong; Feng, Bi-Hong; Huang, Xia; Zhang, Zhi; Sun, Guang-Ming

    2012-06-01

    Polar auxin transport (PAT) plays an important role in the adventitious root formation of mango cotyledon segments, but the molecular mechanism remains unclear. In this study, we cloned a gene encoding an auxin efflux carrier (designated as MiPIN1), and we cloned four genes encoding auxin influx carriers (designated as MiAUX1, MiAUX2, MiAUX3 and MiAUX4). The results of a phylogenetic tree analysis indicated that MiPIN1 and the MiAUXs belong to plant PIN and AUXs/LAXs groups. Quantitative real-time PCR indicated that the expression of MiPIN1 and the MiAUXs was lowest at 0 days but sharply increased on and after day 4. During the root formation in the mango cotyledon segments, the MiPIN1 expression in the distal cut surface (DCS) was always higher than the expression in the proximal cut surface (PCS) whereas the expression of the MiAUXs in the PCS was usually higher than in the DCS. This expression pattern might be result in the PAT from the DCS to the PCS, which is essential for the adventitious root formation in the PCS. Our previous study indicated that a pre-treatment of embryos with indole-3-butyric acid (IBA) significantly promoted adventitious rooting in PCS whereas a pre-treatment with 2,3,5-triiodobenzoic acid (TIBA) completely inhibited this rooting. In this study, however, IBA and TIBA pre-treatments slightly changed the expression of MiPIN1. In contrast, while the MiAUX3 and MiAUX4 expression levels were significantly up-regulated by the IBA pre-treatment, the expression levels were down-regulated by the TIBA pre-treatment. These findings imply that MiAUX3 and MiAUX4 are more sensitive to the IBA and TIBA treatments and that they might play important roles during adventitious root formation in mango cotyledon segments.

  6. Lung vascular cell proliferation in hyperoxic pulmonary hypertension and on return to air: ( sup 3 H)thymidine pulse-labeling of intimal, medial, and adventitial cells in microvessels and at the hilum

    SciTech Connect

    Jones, R.; Adler, C.; Farber, F. )

    1989-11-01

    The labeling index (LI) of lung vascular intimal endothelial and precursor smooth muscle, medial (smooth muscle), and adventitial cells (fibroblasts) was in the normal rat and in the rat with pulmonary hypertension caused by breathing high oxygen (87% O2 at normobaric pressure). Cell labeling was assessed during hyperoxia (Days 1, 4, 7, 10, and 28), at the start and end of weaning hyperoxia-adapted rats to air (Days 29 and 35), and 1, 2, and 4 wk after return to breathing air after hyperoxia and weaning (Days 42, 49, and 63). Bursts of intimal, medial, and adventitial cell proliferation different in their timing and extent contribute to lung vascular wall remodelling in these injuries. The Ll of the microvascular fibroblast increased most on Day 4 of hyperoxia (20-fold); it persisted above the normal rate throughout hyperoxia and was high at the start of weaning (greater than 1- less than 2-fold). Two weeks after return to breathing air, it again increased (less than 1-fold). The Ll of the microvascular endothelial cell increased most on Day 7 of hyperoxia (10-fold); it also persisted above the normal rate throughout hyperoxia and at the start of weaning (greater than 2- less than 3-fold) and increased again 2 wk after return to breathing air (6-fold). Labeled precursor cells were not present in the normal lung: they were present on Days 4 and 7 of hyperoxia but not at any other time, including weaning and return to breathing air.

  7. Adventitial transplantation of blood outgrowth endothelial cells in porcine haemodialysis grafts alleviates hypoxia and decreases neointimal proliferation through a matrix metalloproteinase-9-mediated pathway—a pilot study

    PubMed Central

    Hughes, Deborah; Fu, Alex A.; Puggioni, Alessandra; Glockner, James F.; Anwer, Bilal; McGuire, Antonio M.; Mukhopadhyay, Debabrata; Misra, Sanjay

    2009-01-01

    Purpose. We hypothesized that adventitial transplantation of blood outgrowth endothelial cells (BOEC) to the vein-to-graft anastomosis of polytetrafluoroethylene grafts will reduce neointimal hyperplasia by reducing hypoxia inducible factor-1α (HIF-1α), by increasing angiogenesis in a porcine model of chronic renal insufficiency with haemodialysis polytetrafluoroethylene grafts. Because matrix metalloproteinases (MMPs) have been shown to be involved with angiogenesis, the expression of MMPs and their inhibitors was determined. Methods. Chronic renal insufficiency was created by subtotal renal infarction and 28 days later, arteriovenous PTFE grafts were placed bilaterally from the carotid artery to the jugular vein. Autologous blood outgrowth endothelial cells labeled with Lac Z were transplanted to the adventitia of the vein-to-graft anastomosis using polyglycolic acid scaffolding and scaffolding only to other side (control). Animals were killed 14 days later and vessels were explanted from the vein-to-graft anastomosis of both sides and underwent immunohistochemical analysis, western blotting and zymography for HIF-1α, MMP-2, MMP-9, TIMP-1 and TIMP-2. BOEC were also made hypoxic and normoxic for 12, 24 and 48 h to determine protein expression for MMPs and TIMPs. Results. Under hypoxia, BOEC significantly increased the expression of pro MMP-2 by 12 h and TIMP-2 by 24 h when compared to normoxic cells (P < 0.05). Transplantation of BOEC resulted in a significant decrease in both HIF-1α and intima-to-media ratio with a significant increase in both pro and active MMP-9 when compared to control vessels (P < 0.05). MMP-9 activity was localized to the neointima of the transplanted vessels by immunohistochemistry. There was increased CD31 density with engraftment of BOEC cells into the neointima of both the transplanted vessels compared to controls (P = NS). Conclusion. Transplantation of BOEC resulted in a significant decrease in intimal hyperplasia and HIF-1α with

  8. [Normal and Adventitious Breath Sounds].

    PubMed

    Koehler, U; Hildebrandt, O; Kerzel, S; Urban, C; Hoehle, L; Weissflog, A; Nikolaizik, W; Koehler, J; Sohrabi, K; Gross, V

    2016-06-01

    Auscultation of the lung is an inexpensive, noninvasive and easy-to-perform tool. It is an important part of the physical examination and is help ful to distinguish physiological respiratory sounds from pathophysiological events. Computerized lung sound analysis is a powerful tool for optimizing and quantifying electronic auscultation based on the specific lung sound spectral characteristics. The automatic analysis of respiratory sounds assumes that physiological and pathological sounds are reliably analyzed based on special algorithms. The development of automated long-term lungsound monitors enables objective assessment of different respiratory symptoms.

  9. Effect of Exogenous Indole-3-Acetic Acid and Indole-3-Butyric Acid on Internal Levels of the Respective Auxins and Their Conjugation with Aspartic Acid during Adventitious Root Formation in Pea Cuttings.

    PubMed

    Nordström, A C; Jacobs, F A; Eliasson, L

    1991-07-01

    The influence of exogenous indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) on the internal levels of these auxins was studied during the first 4 days of adventitious root formation in cuttings of Pisum sativum L. The quantitations were done by high performance liquid chromatography with spectrofluorometric detection. IBA, identified by combined gas chromatography-mass spectrometry (GC-MS), was found to naturally occur in this plant material. The root inducing ability of exogenous IBA was superior to that of IAA. The IAA level in the tissue increased considerably on the first day after application of IAA, but rapidly decreased again, returning to a level twice the control by day 3. The predominant metabolic route was conjugation with aspartic acid, as reflected by the increase in the level of indole-3-acetylaspartic acid. The IBA treatment resulted in increases in the levels of IBA, IAA, and indole-3-acetylaspartic acid. The IAA content rapidly returned to control levels, whereas the IBA level remained high throughout the experimental period. High amounts of indole-3-butyrylaspartic acid were found in the tissue after feeding with IBA. The identity of the conjugate was confirmed by (1)H-nuclear magnetic resonance and GC-MS. IBA was much more stable in solution than IAA. No IAA was detected after 48 hours, whereas 70% IBA was still recovered after this time. The relatively higher root inducing ability of IBA is ascribed to the fact that its level remained elevated longer than that of IAA, even though IBA was metabolized in the tissue. Adventitious root formation is discussed on the basis of these findings. PMID:16668265

  10. Use of PCR-based assays for the detection of the adventitious agent porcine circovirus type 1 (PCV1) in vaccines, and for confirming the identity of cell substrates and viruses used in vaccine production.

    PubMed

    Kumar, Deepak; Beach, Nathan M; Meng, Xiang-Jin; Hegde, Nagendra R

    2012-01-01

    Safety and quality are important issues for vaccines. Whereas reversion to virulence poses a safety risk with live attenuated vaccines, the potential for the presence of adventitious agents is also an issue of vaccine quality. The recent detection or porcine circovirus type 1 (PCV1) in human vaccines has further highlighted the importance of quality control in vaccine production. The purpose of this study was to use a novel conventional PCR to detect PCV1, and subsequently screen materials used in the manufacture of vaccines at Bharat Biotech International Limited, India. The genome or gene fragments of PCV1 were not detected in any of the vaccines and materials tested, including the live attenuated rotavirus vaccine candidate ROTAVAC(®). Further, the identity of the cells and the viruses used as starting materials in the manufacture of these vaccines was confirmed by species-specific PCR or virus-specific RT-PCR, and no cross-contamination was detected in any case. The methods can be applied for regular in-house quality control screening of raw materials and seeds/banks, as well as formulated vaccines.

  11. Adventitious bud regeneration from leaf expiants of the shrubby ornamental honeysuckle, Lonicera nitida Wils. cv. 'Maigrün': effects of thidiazuron and 2,3,5-triiodobenzoic acid.

    PubMed

    Cambecèdes, J; Duron, M; Decourtye, L

    1991-11-01

    Different combinations of auxins and cytokinins were employed to assess the regeneration capacity from in vitro leaf explants of Lonicera nitida Wils. cv 'Maïgrün'. A high frequency of rhizogenesis was noticed, with 2.3 μM thidiazuron plus 2.9 μM indole-3-acetic acid as the only hormonal combination to support caulogenic responses. Increasing thidiazuron concentration and/or suppressing auxin did not improve caulogenesis. Combining thidiazuron with 2,3,5-triiodobenzoic acid produced a dramatic increase in the percentage of caulogenic explants. A maximum of 74% of adventitious bud forming explants was obtained with 2.3 μM thidiazuron plus 20 μM 2,3,5-triiodobenzoic acid. Buds were often in a rosette form and were vitreous, so that shoot elongation was difficult to obtain. The effect of the duration of the 2,3,5-triiodobenzoic acid treatment on shoot elongation was investigated. PMID:24221854

  12. Decellularized Allogeneic Heart Valves Demonstrate Self-Regeneration Potential after a Long-Term Preclinical Evaluation

    PubMed Central

    Iop, Laura; Bonetti, Antonella; Naso, Filippo; Rizzo, Stefania; Cagnin, Stefano; Bianco, Roberto; Lin, Carlo Dal; Martini, Paolo; Poser, Helen; Franci, Paolo; Lanfranchi, Gerolamo; Busetto, Roberto; Spina, Michel; Basso, Cristina; Marchini, Maurizio; Gandaglia, Alessandro

    2014-01-01

    Tissue-engineered heart valves are proposed as novel viable replacements granting longer durability and growth potential. However, they require extensive in vitro cell-conditioning in bioreactor before implantation. Here, the propensity of non-preconditioned decellularized heart valves to spontaneous in body self-regeneration was investigated in a large animal model. Decellularized porcine aortic valves were evaluated for right ventricular outflow tract (RVOT) reconstruction in Vietnamese Pigs (n = 11) with 6 (n = 5) and 15 (n = 6) follow-up months. Repositioned native valves (n = 2 for each time) were considered as control. Tissue and cell components from explanted valves were investigated by histology, immunohistochemistry, electron microscopy, and gene expression. Most substitutes constantly demonstrated in vivo adequate hemodynamic performances and ex vivo progressive repopulation during the 15 implantation months without signs of calcifications, fibrosis and/or thrombosis, as revealed by histological, immunohistochemical, ultrastructural, metabolic and transcriptomic profiles. Colonizing cells displayed native-like phenotypes and actively synthesized novel extracellular matrix elements, as collagen and elastin fibers. New mature blood vessels, i.e. capillaries and vasa vasorum, were identified in repopulated valves especially in the medial and adventitial tunicae of regenerated arterial walls. Such findings correlated to the up-regulated vascular gene transcription. Neoinnervation hallmarks were appreciated at histological and ultrastructural levels. Macrophage populations with reparative M2 phenotype were highly represented in repopulated valves. Indeed, no aspects of adverse/immune reaction were revealed in immunohistochemical and transcriptomic patterns. Among differentiated elements, several cells were identified expressing typical stem cell markers of embryonic, hematopoietic, neural and mesenchymal lineages in significantly higher number

  13. Decellularized allogeneic heart valves demonstrate self-regeneration potential after a long-term preclinical evaluation.

    PubMed

    Iop, Laura; Bonetti, Antonella; Naso, Filippo; Rizzo, Stefania; Cagnin, Stefano; Bianco, Roberto; Dal Lin, Carlo; Martini, Paolo; Poser, Helen; Franci, Paolo; Lanfranchi, Gerolamo; Busetto, Roberto; Spina, Michel; Basso, Cristina; Marchini, Maurizio; Gandaglia, Alessandro; Ortolani, Fulvia; Gerosa, Gino

    2014-01-01

    Tissue-engineered heart valves are proposed as novel viable replacements granting longer durability and growth potential. However, they require extensive in vitro cell-conditioning in bioreactor before implantation. Here, the propensity of non-preconditioned decellularized heart valves to spontaneous in body self-regeneration was investigated in a large animal model. Decellularized porcine aortic valves were evaluated for right ventricular outflow tract (RVOT) reconstruction in Vietnamese Pigs (n = 11) with 6 (n = 5) and 15 (n = 6) follow-up months. Repositioned native valves (n = 2 for each time) were considered as control. Tissue and cell components from explanted valves were investigated by histology, immunohistochemistry, electron microscopy, and gene expression. Most substitutes constantly demonstrated in vivo adequate hemodynamic performances and ex vivo progressive repopulation during the 15 implantation months without signs of calcifications, fibrosis and/or thrombosis, as revealed by histological, immunohistochemical, ultrastructural, metabolic and transcriptomic profiles. Colonizing cells displayed native-like phenotypes and actively synthesized novel extracellular matrix elements, as collagen and elastin fibers. New mature blood vessels, i.e. capillaries and vasa vasorum, were identified in repopulated valves especially in the medial and adventitial tunicae of regenerated arterial walls. Such findings correlated to the up-regulated vascular gene transcription. Neoinnervation hallmarks were appreciated at histological and ultrastructural levels. Macrophage populations with reparative M2 phenotype were highly represented in repopulated valves. Indeed, no aspects of adverse/immune reaction were revealed in immunohistochemical and transcriptomic patterns. Among differentiated elements, several cells were identified expressing typical stem cell markers of embryonic, hematopoietic, neural and mesenchymal lineages in significantly higher number

  14. Arabidopsis SHR and SCR transcription factors and AUX1 auxin influx carrier control the switch between adventitious rooting and xylogenesis in planta and in in vitro cultured thin cell layers

    PubMed Central

    Della Rovere, F.; Fattorini, L.; D’Angeli, S.; Veloccia, A.; Del Duca, S.; Cai, G.; Falasca, G.; Altamura, M. M.

    2015-01-01

    Background and Aims Adventitious roots (ARs) are essential for vegetative propagation. The Arabidopsis thaliana transcription factors SHORT ROOT (SHR) and SCARECROW (SCR) affect primary/lateral root development, but their involvement in AR formation is uncertain. LAX3 and AUX1 auxin influx carriers contribute to primary/lateral root development. LAX3 expression is regulated by SHR, and LAX3 contributes to AR tip auxin maximum. In contrast, AUX1 involvement in AR development is unknown. Xylogenesis is induced by auxin plus cytokinin as is AR formation, but the genes involved are largely unknown. Stem thin cell layers (TCLs) form ARs and undergo xylogenesis under the same auxin plus cytokinin input. The aim of this research was to investigate SHR, SCR, AUX1 and LAX3 involvement in AR formation and xylogenesis in intact hypocotyls and stem TCLs in arabidopsis. Methods Hypocotyls of scr-1, shr-1, lax3, aux1-21 and lax3/aux1-21 Arabidopsis thaliana null mutant seedlings grown with or without auxin plus cytokinin were examined histologically, as were stem TCLs cultured with auxin plus cytokinin. SCR and AUX1 expression was monitored using pSCR::GFP and AUX1::GUS lines, and LAX3 expression and auxin localization during xylogenesis were monitored by using LAX3::GUS and DR5::GUS lines. Key Results AR formation was inhibited in all mutants, except lax3. SCR was expressed in pericycle anticlinally derived AR-forming cells of intact hypocotyls, and in cell clumps forming AR meristemoids of TCLs. The apex was anomalous in shr and scr ARs. In all mutant hypocotyls, the pericycle divided periclinally to produce xylogenesis. Xylary element maturation was favoured by auxin plus cytokinin in shr and aux1-21. Xylogenesis was enhanced in TCLs, and in aux1-21 and shr in particular. AUX1 was expressed before LAX3, i.e. in the early derivatives leading to either ARs or xylogenesis. Conclusions AR formation and xylogenesis are developmental programmes that are inversely related, but they

  15. Design factors of intravascular dual frequency transducers for super-harmonic contrast imaging and acoustic angiography

    NASA Astrophysics Data System (ADS)

    Ma, Jianguo; Martin, K. Heath; Li, Yang; Dayton, Paul A.; Shung, K. Kirk; Zhou, Qifa; Jiang, Xiaoning

    2015-05-01

    Imaging of coronary vasa vasorum may lead to assessment of the vulnerable plaque development in diagnosis of atherosclerosis diseases. Dual frequency transducers capable of detection of microbubble super-harmonics have shown promise as a new contrast-enhanced intravascular ultrasound (CE-IVUS) platform with the capability of vasa vasorum imaging. Contrast-to-tissue ratio (CTR) in CE-IVUS imaging can be closely associated with low frequency transmitter performance. In this paper, transducer designs encompassing different transducer layouts, transmitting frequencies, and transducer materials are compared for optimization of imaging performance. In the layout selection, the stacked configuration showed superior super-harmonic imaging compared with the interleaved configuration. In the transmitter frequency selection, a decrease in frequency from 6.5 MHz to 5 MHz resulted in an increase of CTR from 15 dB to 22 dB when receiving frequency was kept constant at 30 MHz. In the material selection, the dual frequency transducer with the lead magnesium niobate-lead titanate (PMN-PT) 1-3 composite transmitter yielded higher axial resolution compared to single crystal transmitters (70 μm compared to 150 μm pulse length). These comparisons provide guidelines for the design of intravascular acoustic angiography transducers.

  16. Design factors of intravascular dual frequency transducers for super-harmonic contrast imaging and acoustic angiography

    PubMed Central

    Ma, Jianguo; Martin, K. Heath; Li, Yang; Dayton, Paul A.; Shung, K. Kirk; Zhou, Qifa; Jiang, Xiaoning

    2015-01-01

    Imaging of coronary vasa vasorum may lead to assessment of the vulnerable plaque development in diagnosis of atherosclerosis diseases. Dual frequency transducers capable of detection of microbubble super-harmonics have shown promise as a new contrast-enhanced intravascular ultrasound (CE-IVUS) platform with the capability of vasa vasorum imaging. Contrast-to-tissue ratio (CTR) in CE-IVUS imaging can be closely associated with the low frequency transmitter performance. In this paper, transducer designs encompassing different transducer layouts, transmitting frequencies, and transducer materials are compared for optimization of imaging performance. In the layout selection, the stacked configuration showed superior super-harmonic imaging compared with the interleaved configuration. In the transmitter frequency selection, a decrease in frequency from 6.5 MHz to 5 MHz resulted in an increase of CTR from 15 dB to 22 dB when receiving frequency was kept constant at 30 MHz. In the material selection, the dual frequency transducer with the lead magnesium niobate-lead titanate (PMN-PT) 1-3 composite transmitter yielded higher axial resolution compared to single crystal transmitters (70 μm compared to 150 μm pulse length). These comparisons provide guidelines for design of intravascular acoustic angiography transducers. PMID:25856384

  17. Microvasular and macrovascular complications in diabetes mellitus: Distinct or continuum?

    PubMed Central

    Chawla, Aastha; Chawla, Rajeev; Jaggi, Shalini

    2016-01-01

    Diabetes and related complications are associated with long-term damage and failure of various organ systems. The line of demarcation between the pathogenic mechanisms of microvascular and macrovascular complications of diabetes and differing responses to therapeutic interventions is blurred. Diabetes induces changes in the microvasculature, causing extracellular matrix protein synthesis, and capillary basement membrane thickening which are the pathognomic features of diabetic microangiopathy. These changes in conjunction with advanced glycation end products, oxidative stress, low grade inflammation, and neovascularization of vasa vasorum can lead to macrovascular complications. Hyperglycemia is the principal cause of microvasculopathy but also appears to play an important role in causation of macrovasculopathy. There is thought to be an intersection between micro and macro vascular complications, but the two disorders seem to be strongly interconnected, with micro vascular diseases promoting atherosclerosis through processes such as hypoxia and changes in vasa vasorum. It is thus imperative to understand whether microvascular complications distinctly precede macrovascular complications or do both of them progress simultaneously as a continuum. This will allow re-focusing on the clinical issues with a unifying perspective which can improve type 2 diabetes mellitus outcomes. PMID:27366724

  18. CONTRAST-ENHANCED INTRAVASCULAR ULTRASOUND PULSE SEQUENCES FOR BANDWIDTH-LIMITED TRANSDUCERS

    PubMed Central

    Maresca, David; Renaud, Guillaume; van Soest, Gijs; Li, Xiang; Zhou, Qifa; Shung, K. Kirk; de Jong, Nico; van der Steen, Antonius F. W.

    2013-01-01

    We demonstrate two methods for vasa vasorum imaging using contrast-enhanced intravascular ultrasound, which can be performed using commercial catheters. Plaque neovascularization was recognized as an independent marker of coronary artery plaque vulnerability. IVUS-based methods to image the microvessels available to date require high bandwidth (−6 dB relative frequency bandwidth >70%), which are not routinely available commercially. We explored the potential of ultraharmonic imaging and chirp reversal imaging for vasa vasorum imaging. In vitro recordings were performed on a tissue-mimicking phantom using a commercial ultrasound contrast agent and a transducer with a center frequency of 34 MHz and a −6 dB relative bandwidth of 56%. Acoustic peak pressures <500 kPa were used. A tissue-mimicking phantom with channels down to 200 μm in diameter was successfully imaged by the two contrast detection sequences while the smallest channel stayed invisible in conventional intravascular ultrasound images. Ultraharmonic imaging provided the best contrast agent detection. PMID:23384459

  19. Design factors of intravascular dual frequency transducers for super-harmonic contrast imaging and acoustic angiography.

    PubMed

    Ma, Jianguo; Martin, K Heath; Li, Yang; Dayton, Paul A; Shung, K Kirk; Zhou, Qifa; Jiang, Xiaoning

    2015-05-01

    Imaging of coronary vasa vasorum may lead to assessment of the vulnerable plaque development in diagnosis of atherosclerosis diseases. Dual frequency transducers capable of detection of microbubble super-harmonics have shown promise as a new contrast-enhanced intravascular ultrasound (CE-IVUS) platform with the capability of vasa vasorum imaging. Contrast-to-tissue ratio (CTR) in CE-IVUS imaging can be closely associated with low frequency transmitter performance. In this paper, transducer designs encompassing different transducer layouts, transmitting frequencies, and transducer materials are compared for optimization of imaging performance. In the layout selection, the stacked configuration showed superior super-harmonic imaging compared with the interleaved configuration. In the transmitter frequency selection, a decrease in frequency from 6.5 MHz to 5 MHz resulted in an increase of CTR from 15 dB to 22 dB when receiving frequency was kept constant at 30 MHz. In the material selection, the dual frequency transducer with the lead magnesium niobate-lead titanate (PMN-PT) 1-3 composite transmitter yielded higher axial resolution compared to single crystal transmitters (70 μm compared to 150 μm pulse length). These comparisons provide guidelines for the design of intravascular acoustic angiography transducers. PMID:25856384

  20. Mutations in smooth muscle alpha-actin (ACTA2) lead to thoracic aortic aneurysms and dissections.

    PubMed

    Guo, Dong-Chuan; Pannu, Hariyadarshi; Tran-Fadulu, Van; Papke, Christina L; Yu, Robert K; Avidan, Nili; Bourgeois, Scott; Estrera, Anthony L; Safi, Hazim J; Sparks, Elizabeth; Amor, David; Ades, Lesley; McConnell, Vivienne; Willoughby, Colin E; Abuelo, Dianne; Willing, Marcia; Lewis, Richard A; Kim, Dong H; Scherer, Steve; Tung, Poyee P; Ahn, Chul; Buja, L Maximilian; Raman, C S; Shete, Sanjay S; Milewicz, Dianna M

    2007-12-01

    The major function of vascular smooth muscle cells (SMCs) is contraction to regulate blood pressure and flow. SMC contractile force requires cyclic interactions between SMC alpha-actin (encoded by ACTA2) and the beta-myosin heavy chain (encoded by MYH11). Here we show that missense mutations in ACTA2 are responsible for 14% of inherited ascending thoracic aortic aneurysms and dissections (TAAD). Structural analyses and immunofluorescence of actin filaments in SMCs derived from individuals heterozygous for ACTA2 mutations illustrate that these mutations interfere with actin filament assembly and are predicted to decrease SMC contraction. Aortic tissues from affected individuals showed aortic medial degeneration, focal areas of medial SMC hyperplasia and disarray, and stenotic arteries in the vasa vasorum due to medial SMC proliferation. These data, along with the previously reported MYH11 mutations causing familial TAAD, indicate the importance of SMC contraction in maintaining the structural integrity of the ascending aorta.

  1. Imaging Atherosclerosis and Vulnerable Plaque

    PubMed Central

    Sadeghi, Mehran M.; Glover, David K.; Lanza, Gregory M.; Fayad, Zahi A.; Johnson, Lynne L.

    2010-01-01

    Identifying patients at high risk for an acute cardiovascular event such as myocardial infarction or stroke and assessing the total atherosclerotic burden are clinically important. Currently available imaging modalities can delineate vascular wall anatomy and, with novel probes, target biologic processes important in plaque evolution and plaque stability. Expansion of the vessel wall involving remodeling of the extracellular matrix can be imaged, as can angiogenesis of the vasa vasorum, plaque inflammation, and fibrin deposits on early nonocclusive vascular thrombosis. Several imaging platforms are available for targeted vascular imaging to acquire information on both anatomy and pathobiology in the same imaging session using either hybrid technology (nuclear combined with CT) or MRI combined with novel probes targeting processes identified by molecular biology to be of importance. This article will discuss the current state of the art of these modalities and challenges to clinical translation. PMID:20395341

  2. A study of the ultrastructure of developing human umbilical vessels.

    PubMed Central

    Sexton, A J; Turmaine, M; Cai, W Q; Burnstock, G

    1996-01-01

    Electron microscopic techniques were used to examine the ultrastructure of developing human umbilical arteries and vein (8-12, 13-17 and 37-40 wk gestational age). These showed that with increasing age there is (1) an increase in the size of the lumen and the thickness of the media; (2) an increase in the ratio of contractile smooth muscle phenotypic cells; (3) an increase in the myofilament content of the smooth muscle cells and the number of Weibel-Palade bodies; (4) a decrease in the glycogen content; (5) an appearance of microvilli on the luminal surface of the endothelium. Lipid vesicles, nerves and vasa vasorum were not observed in any region of the umbilical vein or arteries. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:8655418

  3. The affinity of troxerutin for the venous wall measured by laser scanning microscopy.

    PubMed

    Patwardhan, A; Carlsson, K; Poullain, J C; Taccoen, A; Gerentes, I

    1995-08-01

    The uptake and localization of troxerutin, a trihydroxy-ethyl-rutoside, in the venous wall have been studied in patients undergoing long saphenous vein surgery. Troxerutin, an autofluorescent drug, is currently used to relieve oedema and subjective symptoms in patients with chronic venous insufficiency. In order to determine the localization of the troxerutin, a confocal scanning laser microscope has been used to record the fluorescence from vein cross sections. The quantified fluorescence was used as a measure of the local concentration of troxerutin. In order to reduce the effects of local variation, several images have been scanned from each specimen. Then the recorded data have been analysed to see how the fluorescence varies in the radial direction within the venous wall. Results showed that troxerutin was significantly accumulated in both inner and outer parts of the venous wall. Whereas inner wall troxerutin uptake resulted from direct diffusion through the lumen, the outer wall uptake proceeded likely from the vasa vasorum circulation.

  4. [Aortic inflammatory lesions in Behçet's disease].

    PubMed

    Desbois, A-C; Wechsler, B; Cacoub, P; Saadoun, D

    2016-04-01

    The arterial lesions affect about 10% of patients with Behçet's disease (BD). Aortic inflammatory involvement includes predominantly aortic aneurysmal lesions affecting most often the abdominal aorta. They account for the severity of the disease and are a leading cause of death when they hit the aorta or pulmonary arteries. Within the arterial lesions of BD, aortic involvement is, with femoral lesions, the most common site involved (18-28% of patients with vascular disease). Unlike other large vessels vasculitis (i.e. giant cell arteritis and Takayasu's arteritis) diffuse aortitis is observed in less than 5% of patients with BD. Aortic lesions of BD may be asymptomatic (systematic imaging or occasionally associated with other vascular event) or be revealed by the occurrence of abdominal, thoracic or lumbar pain, or an aortic valve insufficiency. Fever is frequently associated. Increase in acute phase reactants is common in these patients. Histological analysis may show infiltration by lymphocytes, neutrophils and plasma cells in the media and adventitia and a proliferation of the vasa vasorum in the media as well as a fibroblastic proliferation. In the later phase, a fibrous thickening of the media and adventitia is observed as well as a proliferation and thickening of the vasa vasorum. The therapeutic management should always include a medical treatment for the control of inflammation (corticosteroids, immunosuppressive drugs and/or biotherapy) and often an endovascular or surgical treatment if the aneurysm is threatening. The choice between endovascular or surgical treatment is considered case by case, depending on the experience of the team, anatomical conditions and of the clinical presentation. In this review, we provide a detailed and updated review of the literature to describe the aortic inflammatory damage associated with Behçet's disease. PMID:26611428

  5. Inhibition of cyclooxygenase isoforms in late- but not midgestation decreases contractility of the ductus arteriosus and prevents postnatal closure in mice

    PubMed Central

    Reese, Jeff; Anderson, Judy D.; Brown, Naoko; Roman, Christine; Clyman, Ronald I.

    2010-01-01

    Use of cyclooxygenase (COX) inhibitors to delay preterm birth is complicated by in utero constriction of the ductus arteriosus and delayed postnatal closure. Delayed postnatal closure has been attributed to loss of vasa vasorum flow and ductus wall ischemia resulting from constriction in utero. We used the murine ductus (which does not depend on vasa vasorum flow) to determine whether delayed postnatal closure may be because of mechanisms independent of in utero constriction. Acute inhibition of both COX isoforms constricted the fetal ductus on days 18 and 19 (term) but not earlier in gestation; COX-2 inhibition constricted the fetal ductus more than COX-1 inhibition. In contrast, mice exposed to prolonged inhibition of COX-1, COX-2, or both COX isoforms (starting on day 15, when the ductus does not respond to the inhibitors) had no contractile response to the inhibitors on days 18 or 19. Newborn mice closed their ductus within 4 h of birth. Prolonged COX inhibition on days 11–14 of gestation had no effect on newborn ductal closure; however, prolonged COX inhibition on days 15–19 resulted in delayed ductus closure despite exposure to 80% oxygen after birth. Similarly, targeted deletion of COX-2 alone, or COX-1/COX-2 together, impaired postnatal ductus closure. Nitric oxide inhibition did not prevent the delay in ductus closure. These data show that impaired postnatal ductus closure is not the result of in utero ductus constriction or upregulation of nitric oxide synthesis. They are consistent with a novel role for prostaglandins in ductus arteriosus contractile development. PMID:16857891

  6. [Transport and localization of troxerutin in the venous wall].

    PubMed

    Carlsson, K; Patwardhan, A; Poullain, J C; Gerentes, I

    1996-01-01

    The vein wall is nourished by diffusion of blood nutrients from the lumen and from the vasa vasorum. It is likely that drugs take the same ways to reach and diffuse through the vessel wall. Thus the uptake of a drug with affinity for the vein wall should give information on its transport to the tissue. This study aimed to explore troxerutin uptake by the long saphenous vein. Troxerutin is a naturally fluorescent flavonoid which has been known to improve subjective signs of patients with chronic venous insufficiency. Nine patients undergoing surgical treatment of varicosis were enrolled in the study. They received for the last 4 days before surgery either 3,500 mg of troxerutin (n = 4) or 1,000 mg twice daily (n = 2). Three patients as controls did not receive any drug. Two samples from thigh and calf long saphenous vein were harvested in each patients and investigated with a confocal laser scanning microscope developed by our institute measuring the fluorescence emitted by troxerutin after excitation by a 458 nm wavelength laser-beam. The intensity of the overall fluorescence was significantly higher in the treated groups (p < 0,001) and slightly higher in the patient who received 3,500 mg of troxerutin than with the lower dosage. The outer wall region provided the highest fluorescence in the treated group while a significant difference was observed in the fluorescence of the medial region between treated and control group. These results showed a marked affinity of troxerutin for the venous wall. The highest uptake in the outer wall region is likely to result from transport through vasa vasorum, owing to the rheologic properties of the drug. The significant medial fluorescence may account for the venous tone improvement with the drug.

  7. Abdominal aortic aneurysm and histological, clinical, radiological correlation.

    PubMed

    Rodella, Luigi Fabrizio; Rezzani, Rita; Bonomini, Francesca; Peroni, Michele; Cocchi, Marco Angelo; Hirtler, Lena; Bonardelli, Stefano

    2016-04-01

    To date, the pathogenesis of abdominal aortic aneurism (AAA) still remains unclear. As such, the aim of this study was to evaluate changes of the aortic structure during AAA. We analysed the microscopic frame of vessels sections, starting from the primum movens leading to abnormal dilatation. AAA samples were collected and processed through various staining methods (Verhoeff-Van Gieson, Masson Goldner, Sirius Red). Subsequently, the vessel morphology and collagenic web of the tunica media and adventitia were determined and the amount of type I and type III collagen was measured. We also applied immune-histochemistry markers for CD34 and PGP 9.5 in order to identify vascular and nerve structures in the aorta. Immune-positivity quantification was used to calculate the percentage of the stained area. We found increasing deposition of type I collagen and reduced type III collagen in both tunica media and adventitia of AAA. The total amount of vasa vasorum, marked with CD34, and nerva vasorum, marked with PGP 9.5, was also higher in AAA samples. Cardiovascular risk factors (blood pressure, dyslipidemia, cigarette smoking) and radiological data (maximum aneurism diameter, intra-luminal thrombus, aortic wall calcification) increased these changes. These results suggest that the tunica adventitia may have a central role in the pathogenesis of AAA as clearly there are major changes characterized by rooted inflammatory infiltration. The presence of immune components could explain these modifications within the framework of the aorta. PMID:26858185

  8. Stimulation of adventitious rooting of Taxus species by thiamine.

    PubMed

    Chee, P P

    1995-10-01

    Results obtained from using root inducing compounds on Taxus species cuttings suggested that rooting could be significantly enhanced by the presence of thiamine. This observation was verified using a root inducing solution containing a set concentration of IBA (0.2%), NAA (0.1%), and supplemented with various concentrations of thiamine. The best rooting response for Taxus cuspidata stem cuttings was found using this solution supplemented with 0.08% thiamine. Rooted cuttings were easily established and developed into vigorous plants. In addition, Taxus brevifolia shoots obtained from tissue cultures via in vitro organogenesis also responded favorably to this 0.08% thiamine supplemented rooting solution. PMID:24186706

  9. Pathophysiology of diabetic erectile dysfunction: potential contribution of vasa nervorum and advanced glycation endproducts.

    PubMed

    Cellek, S; Cameron, N E; Cotter, M A; Muneer, A

    2013-01-01

    Erectile dysfunction (ED) due to diabetes mellitus remains difficult to treat medically despite advances in pharmacotherapeutic approaches in the field. This unmet need has resulted in a recent re-focus on the pathophysiology, in order to understand the cellular and molecular mechanisms leading to ED in diabetes. Diabetes-induced ED is often resistant to PDE5 inhibitor treatment, thus there is a need to discover targets that may lead to novel approaches for a successful treatment. The aim of this brief review is to update the reader in some of the latest development on that front, with a particular focus on the role of impaired neuronal blood flow and the formation of advanced glycation endproducts.

  10. Pathophysiology of diabetic erectile dysfunction: potential contribution of vasa nervorum and advanced glycation endproducts.

    PubMed

    Cellek, S; Cameron, N E; Cotter, M A; Muneer, A

    2013-01-01

    Erectile dysfunction (ED) due to diabetes mellitus remains difficult to treat medically despite advances in pharmacotherapeutic approaches in the field. This unmet need has resulted in a recent re-focus on the pathophysiology, in order to understand the cellular and molecular mechanisms leading to ED in diabetes. Diabetes-induced ED is often resistant to PDE5 inhibitor treatment, thus there is a need to discover targets that may lead to novel approaches for a successful treatment. The aim of this brief review is to update the reader in some of the latest development on that front, with a particular focus on the role of impaired neuronal blood flow and the formation of advanced glycation endproducts. PMID:22914567

  11. Plaque Composition and No-Reflow Phenomenon During Percutaneous Coronary Intervention of Low-Echoic Structures in Grayscale Intravascular Ultrasound.

    PubMed

    Amano, Hideo; Ikeda, Takanori; Toda, Mikihito; Okubo, Ryo; Yabe, Takayuki; Watanabe, Ippei; Saito, Daiga

    2016-05-25

    It has been reported that coronary vasa vasorum is associated with plaque vulnerability, and low-echoic structures in grayscale intravascular ultrasound (IVUS) are consistent pathologically with vasa vasorum. However, the association of low-echoic structures with plaque composition and no-reflow phenomenon during percutaneous coronary intervention (PCI) is unclear. We investigated plaque composition in virtual histology IVUS (VH-IVUS) and no-reflow phenomenon during PCI of low-echoic structures.A total of 106 lesions being treated by VH-IVUS before PCI were included in this study. Low-echoic structure was defined as a small tubular structure exterior to media without a connection to the vessel lumen in ≥ 3 consecutive crosssectional IVUS images. Lesions with low-echoic structures were found in 42% (45/106).Lesions with low-echoic structures were more prevalent in acute coronary syndrome (ACS) patients (53% [24/45] versus 20% [12/61], P < 0.001), had more positive remodeling (49% [22/45] versus 21% [13/61], P = 0.003), a larger number of VH-IVUS derived thin-cap fibroatheromas (VH-TCFAs) (0.64 ± 0.53 versus 0.05 ± 0.22, P < 0.001), more VH-TCFAs with a baseline plaque burden of 70% or more and minimal luminal area of 4.0 mm(2) or less (29% [13/45] versus 2% [1/61], P < 0.001), and more frequent no-reflow phenomenon after stent implantation and more final TIMI flow grade 0/1/2 (38% [17/45] versus 5% [3/61], P < 0.001; 9% [4/45] versus 0% [0/61], P = 0.03) than lesions without low-echo structures.Lesions with low-echoic structures in grayscale IVUS had high plaque vulnerability and were more prevalent in ACS patients, positive remolding, and VH-TCFAs, and they had more frequent no-reflow phenomenon during PCI than lesions without low-echoic structures.

  12. Pericoronary Adipose Tissue as Storage and Supply Site for Oxidized Low-Density Lipoprotein in Human Coronary Plaques

    PubMed Central

    Uchida, Yasumi; Uchida, Yasuto; Shimoyama, Ei; Hiruta, Nobuyuki; Kishimoto, Toshihiko; Watanabe, Soichiro

    2016-01-01

    Objectives It is generally believed that low-density lipoprotein enters the vascular wall from its lumen and oxidized (oxLDL), after which it plays an important role in atherosclerosis. Because voluminous epicardial adipose tissue is a risk factor for coronary events, there is a possibility that the pericoronary adipose tissue (PCAT), which is a part of epicardial adipose tissue, acts as a risk factor by supplying oxLDL to the coronary arterial wall. The present study was performed whether PCAT stores and supplies oxLDL to the coronary wall. Methods Localization of oxLDL in PCAT and its relation to plaque morphology were examined by immunohistochemical techniques in 27 epicardial coronary arteries excised from 9 human autopsy cases. Results OxLDL deposited in all PCAT of the studied cases. The percent (%) incidence of oxLDL in the intima of 25 normal segment, 19 white plaques, 15 yellow plaques without necrotic core (NC) and 10 yellow plaques with NC, was 32, 84, 93 (p<0.05 vs normal segments and yellow plaques with NC), and 30, respectively. OxLDL deposited either in dotted or diffuse pattern. Double immunohistochemical staining revealed that the dotted oxLDL was that contained in CD68(+)-macrophages. The oxLDL-containing macrophages were observed in the interstitial space but not inside of the vasa vasorum, and they traversed PCAT, adventitia, external and internal elastic laminae, suggesting their migration towards the intima. Diffuse oxLDL deposits were observed in 17 preparations, the majority of which were co-localized with the vasa vasorum in outer or in both inner and outer halves of intima, and rarely in the inner half alone. Conclusions The results suggested that PCAT is a supply source of oxLDL to coronary intima and acts as a risk factor for coronary events, that oxLDL increasingly deposits in the intima with plaque growth and decreases after plaque maturation, and therefore molecular therapies targeting the PCAT before plaque growth could be effective

  13. Radical surgical treatment for recurrent giant fusiform thrombosed vertebral artery aneurysm previously coiled

    PubMed Central

    J-O’Shanahan, Aruma; Noda, Kosumo; Tsuboi, Toshiyuki; Ota, Nakao; Kamiyama, Hiroyasu; Tokuda, Sadahisa; Tanikawa, Rokuya

    2016-01-01

    Background: Fusiform aneurysms are rare (<1%) and the underlying pathophysiology is not well known. Endovascular coiling is the standard of treatment; however, a surgical procedure with vascular reconstruction by excluding the pathological segment of the vessel and restoring the blood flow, seems to be the most effective and definitive treatment. Case Description: We report a patient who presented a fusiform vertebral artery aneurysm previously coiled which developed a giant enlargement and a new contralateral fusiform aneurysm. Hemodynamic changes resulting in the formation of contralateral aneurysm might be the result of aneurysm occlusion without revascularization. In addition, continued blood flow to the aneurysmal wall through the vasa vasorum might result in aneurysm recanalization or regrowth. In order to account for these possible sources of complications, we performed a vascular reconstruction with high and low flow bypasses after trapping the aneurysm. Conclusions: We hypothesize that, in this and similar cases, surgical vascular reconstruction should be the first and definitive treatment under experienced cerebrovascular surgeons. PMID:27127714

  14. A preliminary engineering design of intravascular dual-frequency transducers for contrast-enhanced acoustic angiography and molecular imaging.

    PubMed

    Ma, Jianguo; Martin, K; Dayton, Paul A; Jiang, Xiaoning

    2014-05-01

    Current intravascular ultrasound (IVUS) probes are not optimized for contrast detection because of their design for high-frequency fundamental-mode imaging. However, data from transcutaneous contrast imaging suggests the possibility of utilizing contrast ultrasound for molecular imaging or vasa vasorum assessment to further elucidate atherosclerotic plaque deposition. This paper presents the design, fabrication, and characterization of a small-aperture (0.6 × 3 mm) IVUS probe optimized for high-frequency contrast imaging. The design utilizes a dual-frequency (6.5 MHz/30 MHz) transducer arrangement for exciting microbubbles at low frequencies (near their resonance) and detecting their broadband harmonics at high frequencies, minimizing detected tissue backscatter. The prototype probe is able to generate nonlinear microbubble response with more than 1.2 MPa of rarefractional pressure (mechanical index: 0.48) at 6.5 MHz, and is also able to detect microbubble response with a broadband receiving element (center frequency: 30 MHz, -6-dB fractional bandwidth: 58.6%). Nonlinear super-harmonics from microbubbles flowing through a 200-μm-diameter micro-tube were clearly detected with a signal-to-noise ratio higher than 12 dB. Preliminary phantom imaging at the fundamental frequency (30 MHz) and dual-frequency super-harmonic imaging results suggest the promise of small aperture, dual-frequency IVUS transducers for contrast-enhanced IVUS imaging. PMID:24801226

  15. A Preliminary Engineering Design of Intravascular Dual-Frequency Transducers for Contrast-Enhanced Acoustic Angiography and Molecular Imaging

    PubMed Central

    Ma, Jianguo; Martin, K. Heath; Dayton, Paul A.; Jiang, Xiaoning

    2014-01-01

    Current intravascular ultrasound (IVUS) probes are not optimized for contrast detection because of their design for high-frequency fundamental-mode imaging. However, data from transcutaneous contrast imaging suggests the possibility of utilizing contrast ultrasound for molecular imaging or vasa vasorum assessment to further elucidate atherosclerotic plaque deposition. This paper presents the design, fabrication, and characterization of a small-aperture (0.6 × 3 mm) IVUS probe optimized for high-frequency contrast imaging. The design utilizes a dual-frequency (6.5 MHz/30 MHz) transducer arrangement for exciting microbubbles at low frequencies (near their resonance) and detecting their broadband harmonics at high frequencies, minimizing detected tissue backscatter. The prototype probe is able to generate nonlinear microbubble response with more than 1.2 MPa of rarefractional pressure (mechanical index: 0.48) at 6.5 MHz, and is also able to detect microbubble response with a broadband receiving element (center frequency: 30 MHz, −6-dB fractional bandwidth: 58.6%). Nonlinear super-harmonics from microbubbles flowing through a 200-μm-diameter micro-tube were clearly detected with a signal-to-noise ratio higher than 12 dB. Preliminary phantom imaging at the fundamental frequency (30 MHz) and dual-frequency super-harmonic imaging results suggest the promise of small aperture, dual-frequency IVUS transducers for contrast-enhanced IVUS imaging. PMID:24801226

  16. Effect of a renin-system inhibitor on blood-vessel adaptation in spontaneously hypertensive rats.

    PubMed

    Gould, A B; Goodman, S A

    1987-07-01

    We tested the hypothesis that a metabolic error may be the triggering mechanism which leads to blood-vessel hypertrophy and hypertension. Young spontaneously hypertensive rats (SHR) were fed a moderately high salt diet to exacerbate the purported metabolic error. Haematocrit values and rubidium transport were measured as evidence of renal ATP deficiency and blood-vessel adaptation. The renin system was inhibited in two groups of SHR by giving them enalapril to determine whether angiotensin II was involved in blood-vessel adaptation. Spontaneously hypertensive rats fed the moderately high salt diet had higher haematocrit values than normotensive rats fed the same diet or SHR fed Purina rat food, suggesting a renal ATP deficiency. Spontaneously hypertensive rats had higher Na+,K+-ATPase activity in thoracic aorta after 60 min incubation than a similar group given enalapril (P less than 0.001), suggesting blood-vessel adaptation. Possibly, angiotensin II within the vasa vasorum stimulates hypertrophy which, according to the Folkow hypothesis, leads to higher blood pressure, but may concomitantly increase the respiratory chain units which provide ATP for renal function and ion transport.

  17. Diabetic cardiac autonomic neuropathy: Do we have any treatment perspectives?

    PubMed Central

    Serhiyenko, Victoria A; Serhiyenko, Alexandr A

    2015-01-01

    Cardiac autonomic neuropathy (CAN) is a serious and common complication of diabetes mellitus (DM). Despite its relationship to an increased risk of cardiovascular mortality and its association with multiple symptoms and impairments, the significance of CAN has not been fully appreciated. CAN among DM patients is characterized review the latest evidence and own data regarding the treatment and the treatment perspectives for diabetic CAN. Lifestyle modification, intensive glycemic control might prevent development or progression of CAN. Pathogenetic treatment of CAN includes: balanced diet and physical activity; optimization of glycemic control; treatment of dyslipoproteinemia; correction of metabolic abnormalities in myocardium; prevention and treatment of thrombosis; use of aldose reductase inhibitors; dihomo-γ-linolenic acid (DGLA), acetyl-L-carnitine, antioxidants, first of all α-lipoic acid (α-LA), use of long-chain ω-3 and ω-6 polyunsaturated fatty acids (ω-3 and ω-6 PUFAs), vasodilators, fat-soluble vitamin B1, aminoguanidine; substitutive therapy of growth factors, in severe cases-treatment of orthostatic hypotension. The promising methods include research and use of tools that increase blood flow through the vasa vasorum, including prostacyclin analogues, thromboxane A2 blockers and drugs that contribute into strengthening and/or normalization of Na+, K+-ATPase (phosphodiesterase inhibitor), α-LA, DGLA, ω-3 PUFAs, and the simultaneous prescription of α-LA, ω-3 PUFA and DGLA. PMID:25789106

  18. Is human mating adventitious or the result of lawful choice? A twin study of mate selection.

    PubMed

    Lykken, D T; Tellegen, A

    1993-07-01

    Pairs of middle-aged twins and their spouses provided data on 74 mainly psychological variables. Neither spousal similarity nor idiosyncratic criteria could account for specific mate selection in these 738 couples. Of the twins (and their spouses), 547 independently rated their initial attraction to their twin's mate (or to their spouse's twin): Findings suggest that characteristics both of the chooser and the chosen constrain mate selection only weakly. This article proposes that it is romantic infatuation that commonly determines the final choice from a broad field of potential eligibles and that this phenomenon is inherently random, in the same sense as is imprinting in precocial birds. PMID:8355143

  19. Ribosomal frameshifting and transcriptional slippage: From genetic steganography and cryptography to adventitious use.

    PubMed

    Atkins, John F; Loughran, Gary; Bhatt, Pramod R; Firth, Andrew E; Baranov, Pavel V

    2016-09-01

    Genetic decoding is not 'frozen' as was earlier thought, but dynamic. One facet of this is frameshifting that often results in synthesis of a C-terminal region encoded by a new frame. Ribosomal frameshifting is utilized for the synthesis of additional products, for regulatory purposes and for translational 'correction' of problem or 'savior' indels. Utilization for synthesis of additional products occurs prominently in the decoding of mobile chromosomal element and viral genomes. One class of regulatory frameshifting of stable chromosomal genes governs cellular polyamine levels from yeasts to humans. In many cases of productively utilized frameshifting, the proportion of ribosomes that frameshift at a shift-prone site is enhanced by specific nascent peptide or mRNA context features. Such mRNA signals, which can be 5' or 3' of the shift site or both, can act by pairing with ribosomal RNA or as stem loops or pseudoknots even with one component being 4 kb 3' from the shift site. Transcriptional realignment at slippage-prone sequences also generates productively utilized products encoded trans-frame with respect to the genomic sequence. This too can be enhanced by nucleic acid structure. Together with dynamic codon redefinition, frameshifting is one of the forms of recoding that enriches gene expression.

  20. Transcriptome analysis identifies genes involved in adventitious branches formation of Gracilaria lichenoides in vitro

    PubMed Central

    Wang, Wenlei; Li, Huanqin; Lin, Xiangzhi; Yang, Shanjun; Wang, Zhaokai; Fang, Baishan

    2015-01-01

    Tissue culture could solve the problems associated with Gracilaria cultivation, including the consistent supply of high-quality seed stock, strain improvement, and efficient mass culture of high-yielding commercial strains. However, STC lags behind that of higher plants because of the paucity of genomic information. Transcriptome analysis and the identification of potential unigenes involved in the formation and regeneration of callus or direct induction of ABs are essential. Herein, the CK, EWAB and NPA G. lichenoides transcriptomes were analyzed using the Illumina sequencing platform in first time. A total of 17,922,453,300 nucleotide clean bases were generated and assembled into 21,294 unigenes, providing a total gene space of 400,912,038 nucleotides with an average length of 1,883 and N 50 of 5,055 nucleotides and a G + C content of 52.02%. BLAST analysis resulted in the assignment of 13,724 (97.5%), 3,740 (26.6%), 9,934 (70.6%), 10,611 (75.4%), 9,490 (67.4%), and 7,773 (55.2%) unigenes were annotated to the NR, NT, Swiss-Prot, KEGG, COG, and GO databases, respectively, and the total of annotated unigenes was 14,070. A total of 17,099 transcripts were predicted to possess open reading frames, including 3,238 predicted and 13,861 blasted based on protein databases. In addition, 3,287 SSRs were detected in G.lichenoides, providing further support for genetic variation and marker-assisted selection in the future. Our results suggest that auxin polar transport, auxin signal transduction, crosstalk with other endogenous plant hormones and antioxidant systems, play important roles for ABs formation in G. lichenoides explants in vitro. The present findings will facilitate further studies on gene discovery and on the molecular mechanisms underlying the tissue culture of seaweed. PMID:26657019

  1. Organic adlayer on inorganic materials: XPS analysis selectivity to cope with adventitious contamination

    NASA Astrophysics Data System (ADS)

    Landoulsi, Jessem; Genet, Michel J.; Fleith, Sandrine; Touré, Yetioman; Liascukiene, Irma; Méthivier, Christophe; Rouxhet, Paul G.

    2016-10-01

    This work addresses the ubiquitous presence of organic contaminants at inorganic solid surfaces and the improvement of XPS analysis selectivity to cope with it. Water contact angle measurements showed that the adsorption of organic contaminants occurs readily in ambient air, and faster and more extensively under high vacuum. It is stronger on stainless steel (SS) compared to silica and is significantly reduced when SS is sterilized by autoclaving. The reliability of XPS data was evaluated (selectivity, precision, accuracy) by correlations between spectral data incorporating a large amount of results obtained with different XPS spectrometers on SS and glass samples cleaned in different ways and conditioned with several biomacromolecules. The methodology used allows a discrimination to be made between contaminants and deliberately adsorbed biomacromolecules, and offers perspectives for tracking the source of contamination. Furthermore, a discrimination can be made between oxygen from the organic adlayer and oxygen from the substrate, and the O 1s component above 532.0 eV observed for SS is shown to be due to organic contaminants rather than adsorbed water. This approach offers new perspectives to examine the interactions (displacement or not) between contaminants and compounds of interest, e.g. proteins, at the stage of the adsorption process.

  2. Effect of Adventitious Carbon on the Environmental Degradation of SiC/BN/SiC Composites

    NASA Technical Reports Server (NTRS)

    Ogbuji, L. U. J. T.; Yun, H. M.; DiCarlo, J.

    2002-01-01

    Pesting remains a major obstacle to the application of SiC/SiC composites in engine service and selective degradation of the boron nitride interphase at intermediate temperatures is of primary concern. However, significant progress has been made on interphase improvement recently and we now know more about the phenomenon and ways to suppress it. By screening SiC/BN/SiC materials through characterization of strength and microstructures after exposure in a burner rig, some factors that control pesting in these composites have been determined. A key precaution is careful control of elemental carbon presence in the interphase region.

  3. Ribosomal frameshifting and transcriptional slippage: From genetic steganography and cryptography to adventitious use.

    PubMed

    Atkins, John F; Loughran, Gary; Bhatt, Pramod R; Firth, Andrew E; Baranov, Pavel V

    2016-09-01

    Genetic decoding is not 'frozen' as was earlier thought, but dynamic. One facet of this is frameshifting that often results in synthesis of a C-terminal region encoded by a new frame. Ribosomal frameshifting is utilized for the synthesis of additional products, for regulatory purposes and for translational 'correction' of problem or 'savior' indels. Utilization for synthesis of additional products occurs prominently in the decoding of mobile chromosomal element and viral genomes. One class of regulatory frameshifting of stable chromosomal genes governs cellular polyamine levels from yeasts to humans. In many cases of productively utilized frameshifting, the proportion of ribosomes that frameshift at a shift-prone site is enhanced by specific nascent peptide or mRNA context features. Such mRNA signals, which can be 5' or 3' of the shift site or both, can act by pairing with ribosomal RNA or as stem loops or pseudoknots even with one component being 4 kb 3' from the shift site. Transcriptional realignment at slippage-prone sequences also generates productively utilized products encoded trans-frame with respect to the genomic sequence. This too can be enhanced by nucleic acid structure. Together with dynamic codon redefinition, frameshifting is one of the forms of recoding that enriches gene expression. PMID:27436286

  4. Transcriptome analysis identifies genes involved in adventitious branches formation of Gracilaria lichenoides in vitro.

    PubMed

    Wang, Wenlei; Li, Huanqin; Lin, Xiangzhi; Yang, Shanjun; Wang, Zhaokai; Fang, Baishan

    2015-01-01

    Tissue culture could solve the problems associated with Gracilaria cultivation, including the consistent supply of high-quality seed stock, strain improvement, and efficient mass culture of high-yielding commercial strains. However, STC lags behind that of higher plants because of the paucity of genomic information. Transcriptome analysis and the identification of potential unigenes involved in the formation and regeneration of callus or direct induction of ABs are essential. Herein, the CK, EWAB and NPA G. lichenoides transcriptomes were analyzed using the Illumina sequencing platform in first time. A total of 17,922,453,300 nucleotide clean bases were generated and assembled into 21,294 unigenes, providing a total gene space of 400,912,038 nucleotides with an average length of 1,883 and N 50 of 5,055 nucleotides and a G + C content of 52.02%. BLAST analysis resulted in the assignment of 13,724 (97.5%), 3,740 (26.6%), 9,934 (70.6%), 10,611 (75.4%), 9,490 (67.4%), and 7,773 (55.2%) unigenes were annotated to the NR, NT, Swiss-Prot, KEGG, COG, and GO databases, respectively, and the total of annotated unigenes was 14,070. A total of 17,099 transcripts were predicted to possess open reading frames, including 3,238 predicted and 13,861 blasted based on protein databases. In addition, 3,287 SSRs were detected in G.lichenoides, providing further support for genetic variation and marker-assisted selection in the future. Our results suggest that auxin polar transport, auxin signal transduction, crosstalk with other endogenous plant hormones and antioxidant systems, play important roles for ABs formation in G. lichenoides explants in vitro. The present findings will facilitate further studies on gene discovery and on the molecular mechanisms underlying the tissue culture of seaweed. PMID:26657019

  5. IL-4 gene expression in adventitial layer (fibrous layer) of hepatic ovine and bovine hydatid cysts.

    PubMed

    Dorosti, Zahra; Tolouei, Sepideh; Khanahmad, Hossein; Jafari, Rasool; Jafaee, Fereshteh; Sharafi, Seyedeh Marayam; Darani, Hossein Yousofi

    2016-09-01

    Cystic Echinococcosis is a parasitic disease with cosmopolitan distribution caused by the tape worm Echinococcus granulosus. Fibrous layer is developed around the cyst as a host immune response reaction. The aim of this study was to evaluate the rate of IL-4 gene expression in fibrous layer of bovine and ovine hepatic hydatid cysts using quantitative technique of Real-Time PCR. In this descriptive study the samples of hydatid cyst fibrous layer were taken from 6 bovine and 6 ovine hepatic hydatid cysts. Samples of normal liver tissue close to the cyst were also taken as controls. Total RNA from each sample was extracted and then converted to cDNA. Afterward, the rate of IL-4 gene expression for each sample was evaluated using real-time PCR technique. Data were analyzed by REST software (version 2.0.13, 2009). In sheep the rate of IL-4 gene expression in the fibrous layer of hepatic hydatid cysts was 1.98 times more than the rate of IL4 gene expression in control samples, but the difference was not significant (P = 0.561). In cattle the rate of IL-4 gene expression in the fibrous layer of hepatic hydatid cysts was 9.84 times more than that of control samples which was statistically significant (P < 0.001). With high rate of IL4 expression especially in fibrous layer of bovine hydatid cyst, it can be concluded that this interleukin may play an important role in host parasite relationship. PMID:27605798

  6. Transcriptome analysis identifies genes involved in adventitious branches formation of Gracilaria lichenoides in vitro.

    PubMed

    Wang, Wenlei; Li, Huanqin; Lin, Xiangzhi; Yang, Shanjun; Wang, Zhaokai; Fang, Baishan

    2015-12-11

    Tissue culture could solve the problems associated with Gracilaria cultivation, including the consistent supply of high-quality seed stock, strain improvement, and efficient mass culture of high-yielding commercial strains. However, STC lags behind that of higher plants because of the paucity of genomic information. Transcriptome analysis and the identification of potential unigenes involved in the formation and regeneration of callus or direct induction of ABs are essential. Herein, the CK, EWAB and NPA G. lichenoides transcriptomes were analyzed using the Illumina sequencing platform in first time. A total of 17,922,453,300 nucleotide clean bases were generated and assembled into 21,294 unigenes, providing a total gene space of 400,912,038 nucleotides with an average length of 1,883 and N 50 of 5,055 nucleotides and a G + C content of 52.02%. BLAST analysis resulted in the assignment of 13,724 (97.5%), 3,740 (26.6%), 9,934 (70.6%), 10,611 (75.4%), 9,490 (67.4%), and 7,773 (55.2%) unigenes were annotated to the NR, NT, Swiss-Prot, KEGG, COG, and GO databases, respectively, and the total of annotated unigenes was 14,070. A total of 17,099 transcripts were predicted to possess open reading frames, including 3,238 predicted and 13,861 blasted based on protein databases. In addition, 3,287 SSRs were detected in G.lichenoides, providing further support for genetic variation and marker-assisted selection in the future. Our results suggest that auxin polar transport, auxin signal transduction, crosstalk with other endogenous plant hormones and antioxidant systems, play important roles for ABs formation in G. lichenoides explants in vitro. The present findings will facilitate further studies on gene discovery and on the molecular mechanisms underlying the tissue culture of seaweed.

  7. Chronic intrauterine pulmonary hypertension increases main pulmonary artery stiffness and adventitial remodeling in fetal sheep

    PubMed Central

    Morgan, Matthew R.; Galambos, Csaba; Hunter, Kendall S.; Abman, Steven H.

    2014-01-01

    Persistent pulmonary hypertension of the newborn (PPHN) is a clinical syndrome that is characterized by high pulmonary vascular resistance due to changes in lung vascular growth, structure, and tone. PPHN has been primarily considered as a disease of the small pulmonary arteries (PA), but proximal vascular stiffness has been shown to be an important predictor of morbidity and mortality in other diseases associated with pulmonary hypertension (PH). The objective of this study is to characterize main PA (MPA) stiffness in experimental PPHN and to determine the relationship of altered biomechanics of the MPA with changes in extracellular matrix (ECM) content and orientation of collagen and elastin fibers. MPAs were isolated from control and PPHN fetal sheep model and were tested by planar biaxial testing to measure stiffness in circumferential and axial vessel orientations. Test specimens were fixed for histological assessments of the vascular wall ECM constituents collagen and elastin. MPAs from PPHN sheep had increased mechanical stiffness (P < 0.05) and altered ECM remodeling compared with control MPA. A constitutive mathematical model and histology demonstrated that PPHN vessels have a smaller contribution of elastin and a greater role for collagen fiber engagement compared with the control arteries. We conclude that exposure to chronic hemodynamic stress in late-gestation fetal sheep increases proximal PA stiffness and alters ECM remodeling. We speculate that proximal PA stiffness further contributes to increased right ventricular impedance in experimental PPHN, which contributes to abnormal transition of the pulmonary circulation at birth. PMID:25326575

  8. Micropropagation of Codiaeum variegatum (L.) Blume and regeneration induction via adventitious buds and somatic embryogenesis.

    PubMed

    Radice, Silvia

    2010-01-01

    Codiaeum variegatum (L) Blume cv. "Corazon de oro" and cv. "Norma" are successfully micropropagated when culture are initiated with explants taken from newly sprouted shoots. The establishment and multiplication steps are possible when 1 mg/L BA or 1 mg/L IAA and 3 mg/L 2iP are added to MS medium, according to the cultivar respectively selected.Adventive organogenesis and somatic embryogenesis are induced from leaf explants taken from in vitro buds of croton. On leaf-sectioned of "Corazon de oro" cultured in vitro, 1 mg/L BA stimulates continuous somatic embryos development and induces some shoots too. Replacing BA with 1 mg/L TDZ induces up to 100% bud regeneration in the same explants. On the other hand, leaf-sectioned of C. variegatum cv. Norma does not start somatic embryo differentiation if 1 mg/L TDZ is not added to the MS basal medium. Incipient callus is observed after 30 days of culture, and then, subculture to MS with 1 mg/L BA allows the same process to show on the "Corazon de oro" cultivar. Somatic embryos show growth arrest that is partially overcome by transfer to hormone-free basal medium with activated charcoal. Root induction is possible on basal medium plus 1 mg/L IBA. Plantlets in the greenhouse have variegated leaves true-to-type.

  9. Ribosomal frameshifting and transcriptional slippage: From genetic steganography and cryptography to adventitious use

    PubMed Central

    Atkins, John F.; Loughran, Gary; Bhatt, Pramod R.; Firth, Andrew E.; Baranov, Pavel V.

    2016-01-01

    Genetic decoding is not ‘frozen’ as was earlier thought, but dynamic. One facet of this is frameshifting that often results in synthesis of a C-terminal region encoded by a new frame. Ribosomal frameshifting is utilized for the synthesis of additional products, for regulatory purposes and for translational ‘correction’ of problem or ‘savior’ indels. Utilization for synthesis of additional products occurs prominently in the decoding of mobile chromosomal element and viral genomes. One class of regulatory frameshifting of stable chromosomal genes governs cellular polyamine levels from yeasts to humans. In many cases of productively utilized frameshifting, the proportion of ribosomes that frameshift at a shift-prone site is enhanced by specific nascent peptide or mRNA context features. Such mRNA signals, which can be 5′ or 3′ of the shift site or both, can act by pairing with ribosomal RNA or as stem loops or pseudoknots even with one component being 4 kb 3′ from the shift site. Transcriptional realignment at slippage-prone sequences also generates productively utilized products encoded trans-frame with respect to the genomic sequence. This too can be enhanced by nucleic acid structure. Together with dynamic codon redefinition, frameshifting is one of the forms of recoding that enriches gene expression. PMID:27436286

  10. Alfalfa transgene dispersal and adventitious presence: understanding grower perception of risk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recognizing the importance of coexistence, the alfalfa industry has developed a set of Best Management Practices (BMP) to maintain separation of GE and conventional production. But the success of BMP depends upon the degree that growers comply. Therefore we surveyed 530 alfalfa hay and seed producer...

  11. Regulation of angiogenesis, mural cell recruitment and adventitial macrophage behavior by Toll-like receptors.

    PubMed

    Aplin, Alfred C; Ligresti, Giovanni; Fogel, Eric; Zorzi, Penelope; Smith, Kelly; Nicosia, Roberto F

    2014-01-01

    The angiogenic response to injury can be studied by culturing rat or mouse aortic explants in collagen gels. Gene expression studies show that aortic angiogenesis is preceded by an immune reaction with overexpression of Toll-like receptors (TLRs) and TLR-inducible genes. TLR1, 3, and 6 are transiently upregulated at 24 h whereas TLR2, 4, and 8 expression peaks at 24 h but remains elevated during angiogenesis and vascular regression. Expression of TLR5, 7 and 9 steadily increases over time and is highest during vascular regression. Studies with isolated cells show that TLRs are expressed at higher levels in aortic macrophages compared to endothelial or mural cells with the exception of TLR2 and TLR9 which are more abundant in the aortic endothelium. LPS and other TLR ligands dose dependently stimulate angiogenesis and vascular endothelial growth factor production. TLR9 ligands also influence the behavior of nonendothelial cell types by blocking mural cell recruitment and inducing formation of multinucleated giant cells by macrophages. TLR9-induced mural cell depletion is associated with reduced expression of the mural cell recruiting factor PDGFB. The spontaneous angiogenic response of the aortic rings to injury is reduced in cultures from mice deficient in myeloid differentiation primary response 88 (MyD88), a key adapter molecule of TLRs, and following treatment with an inhibitor of the NFκB pathway. These results suggest that the TLR system participates in the angiogenic response of the vessel wall to injury and may play an important role in the regulation of inflammatory angiogenesis in reactive and pathologic processes.

  12. Pressure profile and morphology of the arteries along the giraffe limb.

    PubMed

    Østergaard, Kristine Hovkjaer; Bertelsen, Mads F; Brøndum, Emil T; Aalkjaer, Christian; Hasenkam, J Michael; Smerup, Morten; Wang, Tobias; Nyengaard, Jens Randel; Baandrup, Ulrik

    2011-07-01

    Giraffes are the tallest animals on earth and the effects of gravity on their cardiovascular system have puzzled physiologists for centuries. The authors measured arterial and venous pressure in the foreleg of anesthetized giraffes, suspended in upright standing position, and determined the ratio between tunica media and lumen areas along the length of the femoral/tibial arteries in the hindleg. Volume fraction of elastin, density of vasa vasorum and innervations was estimated by stereology. Immunohistological staining with S100 was used to examine the innervation. The pressure increase in the artery and vein along the foreleg was not significantly different from what was expected on basis of gravity. The area of the arterial lumen in the hindleg decreased towards the hoof from 11.2 ± 4.2 to 0.6 ± 0.5 mm(2) (n = 10, P = 0.001), but most of this narrowing occurred within 2-4 cm immediately below the knee. This abrupt narrowing was associated with a marked increase in media to lumen area ratio (from 1.2 ± 0.5 to 7.8 ± 2.5; P = 0.001), and a decrease in mean volume fraction of elastin from 38 ± 6% proximal to the narrowing to 5.8 ± 1.1% distally (P = 0.001). The narrowing had a six-fold higher innervation density than the immediate distal and proximal regions. The sudden narrowing was also observed in the hind legs of neonates, indicating that it does not develop as an adaptation to the high transmural pressure in the standing giraffe. More likely it represents a preadaptation to the high pressures experienced by adult giraffes.

  13. Vascular ultrasound for atherosclerosis imaging

    PubMed Central

    de Korte, Chris L.; Hansen, Hendrik H. G.; van der Steen, Anton F. W.

    2011-01-01

    Cardiovascular disease is a leading cause of death in the Western world. Therefore, detection and quantification of atherosclerotic disease is of paramount importance to monitor treatment and possible prevention of acute events. Vascular ultrasound is an excellent technique to assess the geometry of vessel walls and plaques. The high temporal as well as spatial resolution allows quantification of luminal area and plaque size and volume. While carotid arteries can be imaged non-invasively, scanning of coronary arteries requires invasive intravascular catheters. Both techniques have already demonstrated their clinical applicability. Using linear array technology, detection of disease as well as monitoring of pharmaceutical treatment in carotid arteries are feasible. Data acquired with intravascular ultrasound catheters have proved to be especially beneficial in understanding the development of atherosclerotic disease in coronary arteries. With the introduction of vascular elastography not only the geometry of plaques but also the risk for rupture of plaques might be identified. These so-called vulnerable plaques are frequently not flow-limiting and rupture of these plaques is responsible for the majority of cerebral and cardiac ischaemic events. Intravascular ultrasound elastography studies have demonstrated a high correlation between high strain and vulnerable plaque features, both ex vivo and in vivo. Additionally, pharmaceutical intervention could be monitored using this technique. Non-invasive vascular elastography has recently been developed for carotid applications by using compound scanning. Validation and initial clinical evaluation is currently being performed. Since abundance of vasa vasorum (VV) is correlated with vulnerable plaque development, quantification of VV might be a unique tool to even prevent this from happening. Using ultrasound contrast agents, it has been demonstrated that VV can be identified and quantified. Although far from routine

  14. The neutral sphingomyelinase-2 is involved in angiogenic signaling triggered by oxidized LDL.

    PubMed

    Camaré, Caroline; Augé, Nathalie; Pucelle, Mélanie; Saint-Lebes, Bertrand; Grazide, Marie-Hélène; Nègre-Salvayre, Anne; Salvayre, Robert

    2016-04-01

    Capillaries of the external part of the normal arterial wall constitute the vasa vasorum network. In atherosclerotic lesions, neovascularization occurs in areas of intimal hyperplasia where it may promote plaque expansion, and intraplaque hemorrhage. Oxidized LDL that are present in atherosclerotic areas activate various angiogenic signaling pathways, including reactive oxygen species and the sphingosine kinase/sphingosine-1-phosphate pathway. We aimed to investigate whether oxidized LDL-induced angiogenesis requires neutral sphingomyelinase-2 activation and the neutral sphingomyelinase-2/sphingosine kinase-1 pathway. The role of neutral sphingomyelinase-2 in angiogenic signaling was investigated in Human Microvascular Endothelial Cells (HMEC-1) forming capillary tube on Matrigel and in vivo in the Matrigel plug assay in C57BL/6 mice and in the chicken chorioallantoic membrane model. Low concentration of human oxidized LDL elicits HMEC-1 capillary tube formation and neutral sphingomyelinase-2 activation, which were blocked by neutral sphingomyelinase-2 inhibitors, GW4869 and specific siRNA. This angiogenic effect was mimicked by low concentration of C6-Ceramide and was inhibited by sphingosine kinase-1 inhibitors. Upstream of neutral sphingomyelinase-2, oxidized LDL-induced activation required LOX-1, reactive oxygen species generation by NADPH oxidase and p38-MAPK activation. Inhibition of sphingosine kinase-1 blocked the angiogenic response and triggered HMEC-1 apoptosis. Low concentration of oxidized LDL was angiogenic in vivo, both in the Matrigel plug assay in mice and in the chorioallantoic membrane model, and was blocked by GW4869. In conclusion, low oxLDL concentration triggers sprouting angiogenesis that involves ROS-induced activation of the neutral sphingomyelinase-2/sphingosine kinase-1 pathway, and is effectively inhibited by GW4869. PMID:26855418

  15. Pressure profile and morphology of the arteries along the giraffe limb.

    PubMed

    Østergaard, Kristine Hovkjaer; Bertelsen, Mads F; Brøndum, Emil T; Aalkjaer, Christian; Hasenkam, J Michael; Smerup, Morten; Wang, Tobias; Nyengaard, Jens Randel; Baandrup, Ulrik

    2011-07-01

    Giraffes are the tallest animals on earth and the effects of gravity on their cardiovascular system have puzzled physiologists for centuries. The authors measured arterial and venous pressure in the foreleg of anesthetized giraffes, suspended in upright standing position, and determined the ratio between tunica media and lumen areas along the length of the femoral/tibial arteries in the hindleg. Volume fraction of elastin, density of vasa vasorum and innervations was estimated by stereology. Immunohistological staining with S100 was used to examine the innervation. The pressure increase in the artery and vein along the foreleg was not significantly different from what was expected on basis of gravity. The area of the arterial lumen in the hindleg decreased towards the hoof from 11.2 ± 4.2 to 0.6 ± 0.5 mm(2) (n = 10, P = 0.001), but most of this narrowing occurred within 2-4 cm immediately below the knee. This abrupt narrowing was associated with a marked increase in media to lumen area ratio (from 1.2 ± 0.5 to 7.8 ± 2.5; P = 0.001), and a decrease in mean volume fraction of elastin from 38 ± 6% proximal to the narrowing to 5.8 ± 1.1% distally (P = 0.001). The narrowing had a six-fold higher innervation density than the immediate distal and proximal regions. The sudden narrowing was also observed in the hind legs of neonates, indicating that it does not develop as an adaptation to the high transmural pressure in the standing giraffe. More likely it represents a preadaptation to the high pressures experienced by adult giraffes. PMID:21207038

  16. Comparative characterization of stromal vascular cells derived from three types of vascular wall and adipose tissue.

    PubMed

    Yang, Santsun; Eto, Hitomi; Kato, Harunosuke; Doi, Kentaro; Kuno, Shinichiro; Kinoshita, Kahori; Ma, Hsu; Tsai, Chi-Han; Chou, Wan-Ting; Yoshimura, Kotaro

    2013-12-01

    Multipotent stem/progenitor cells localize perivascularly in many organs and vessel walls. These tissue-resident stem/progenitor cells differentiate into vascular endothelial cells, pericytes, and other mesenchymal lineages, and participate in physiological maintenance and repair of vasculatures. In this study, we characterized stromal vascular cells obtained through the explant culture method from three different vessel walls in humans: arterial wall (ART; >500 μm in diameter), venous wall (VN; >500 μm in diameter), and small vessels in adipose tissue (SV; arterioles and venules, <100 μm in diameter). These were examined for functionality and compared with adipose-derived stem/stromal cells (ASCs). All stromal vascular cells of different origins presented fibroblast-like morphology and we could not visually discriminate one population from another. Flow cytometry showed that the cultured population heterogeneously expressed a variety of surface antigens associated with stem/progenitor cells, but CD105 was expressed by most cells in all groups, suggesting that the cells generally shared the characteristics of mesenchymal stem cells. Our histological and flow cytometric data suggested that the main population of vessel wall-derived stromal vascular cells were CD34(+)/CD31(-) and came from the tunica adventitia and areola tissue surrounding the adventitia. CD271 (p75NTR) was expressed by the vasa vasorum in the VN adventitia and by a limited population in the adventitia of SV. All three populations differentiated into multiple lineages as did ASCs. ART cells induced the largest quantity of calcium formation in the osteogenic medium, whereas ASCs showed the greatest adipogenic differentiation. SV and VN stromal cells had greater potency for network formation than did ART stromal cells. In conclusion, the three stromal vascular populations exhibited differential functional properties. Our results have clinical implications for vascular diseases such as

  17. Morphological and Molecular Characterization of Human Dermal Lymphatic Collectors

    PubMed Central

    Buttler, Kerstin; Ströbel, Philipp; Becker, Jürgen; Aung, Thiha; Felmerer, Gunther; Wilting, Jörg

    2016-01-01

    Millions of patients suffer from lymphedema worldwide. Supporting the contractility of lymphatic collectors is an attractive target for pharmacological therapy of lymphedema. However, lymphatics have mostly been studied in animals, while the cellular and molecular characteristics of human lymphatic collectors are largely unknown. We studied epifascial lymphatic collectors of the thigh, which were isolated for autologous transplantations. Our immunohistological studies identify additional markers for LECs (vimentin, CCBE1). We show and confirm differences between initial and collecting lymphatics concerning the markers ESAM1, D2-40 and LYVE-1. Our transmission electron microscopic studies reveal two types of smooth muscle cells (SMCs) in the media of the collectors with dark and light cytoplasm. We observed vasa vasorum in the media of the largest collectors, as well as interstitial Cajal-like cells, which are highly ramified cells with long processes, caveolae, and lacking a basal lamina. They are in close contact with SMCs, which possess multiple caveolae at the contact sites. Immunohistologically we identified such cells with antibodies against vimentin and PDGFRα, but not CD34 and cKIT. With Next Generation Sequencing we searched for highly expressed genes in the media of lymphatic collectors, and found therapeutic targets, suitable for acceleration of lymphatic contractility, such as neuropeptide Y receptors 1, and 5; tachykinin receptors 1, and 2; purinergic receptors P2RX1, and 6, P2RY12, 13, and 14; 5-hydroxytryptamine receptors HTR2B, and 3C; and adrenoceptors α2A,B,C. Our studies represent the first comprehensive characterization of human epifascial lymphatic collectors, as a prerequisite for diagnosis and therapy. PMID:27764183

  18. Challenging the Surgical Rodent Hindlimb Ischemia Model with the Miniinterventional Technique

    PubMed Central

    Zhuang, Zhen W.; Shi, Jing; Rhodes, John M.; Tsapakos, Michael J.; Simons, Michael

    2012-01-01

    Purpose To develop an interventional hindlimb ischemic model and compare its angiogenic effect versus surgical ligation (SL) and excision of the femoral artery in rats treated with transplantation of bone marrow mononuclear cells (MNCs) as an angiogenic stimulator. Materials and Methods Forty-eight Lewis rats randomly received interventional embolization (IE) with hydrogel wire or SL and excision of the right femoral artery. Rodents were intraarterially transplanted with 1.5 × 107 MNCs in 500 μL medium from 24 isogenic donor rats. Functional and structural recovery was evaluated by laser Doppler imaging (LDI), cytokine/chemokine assay, and histologic staining. Results In vivo microscopic images showed significantly dilated vasa vasorum around the embolized segment of the right femoral artery at 3 days compared with disorganized tissue structure in the SL group. However, the LDI index was significantly higher in the SL group at 3 days compared with the IE group. LDI did not significantly differ between the two groups at 2 weeks after transplantation. Cytokine assay showed higher levels of interleukin (IL)–1α and IL-18 in the SL group; the IE group had higher levels of interferon-γ, IL-6, IL-13, and granulocyte colony-stimulating factor. Histologic examination demonstrated inflammatory infiltration near the incision within nerve fibers with dilated capillaries, showing nerve degeneration in the SL group. At 2 weeks, histologic analysis demonstrated massive scarring under the skin spreading into the musculature in the SL group. Conclusions A minimally invasive hindlimb ischemia model has been successfully developed that preserves tissue integrity and minimizes inflammation and confounding factors in the early stages of angiogenesis and arteriogenesis. PMID:21459613

  19. Role of Microvascular Tone and Extracellular Matrix Contraction in the Regulation of Interstitial Fluid: Implications for Aortic Dissection.

    PubMed

    Mallat, Ziad; Tedgui, Alain; Henrion, Daniel

    2016-09-01

    The pathophysiology of aortic dissection is poorly understood, and its risk is resistant to medical treatment. Most studies have focused on a proposed pathogenic role of transforming growth factor-β in Marfan disease and related thoracic aortic aneurysms and aortic dissections. However, clinical testing of this concept using angiotensin II type 1 receptor antagonists to block transforming growth factor-β signaling fell short of promise. Genetic mutations that predispose to thoracic aortic aneurysms and aortic dissections affect components of the extracellular matrix and proteins involved in cellular force generation. Thus, a role for dysfunctional mechanosensing in abnormal aortic wall remodeling is emerging. However, how abnormal mechanosensing leads to aortic dissection remains a mystery. Here, we review current knowledge about the regulation of interstitial fluid dynamics and myogenic tone and propose that alteration in contractile force reduces vascular tone in the microcirculation (here, aortic vasa vasorum) and leads to elevations of blood flow, transmural pressure, and fluid flux into the surrounding aortic media. Furthermore, reduced contractile force in medial smooth muscle cells coupled with alteration of structural components of the extracellular matrix limits extracellular matrix contraction, further promoting the formation of intramural edema, a critical step in the initiation of aortic dissection. The concept is supported by several pathophysiological and clinical observations. A direct implication of this concept is that drugs that lower blood pressure and limit interstitial fluid accumulation while preserving or increasing microvascular tone would limit the risk of dissection. In contrast, drugs that substantially lower microvascular tone would be ineffective or may accelerate the disease and precipitate aortic dissection. PMID:27444198

  20. Devascularization of Head and Neck Paragangliomas by Direct Percutaneous Embolization

    SciTech Connect

    Ozyer, Umut Harman, Ali; Yildirim, Erkan; Aytekin, Cuneyt; Akay, Tankut Hakki; Boyvat, Fatih

    2010-10-15

    Preoperative transarterial embolization of head and neck paragangliomas using particulate agents has proven beneficial for decreasing intraoperative blood loss. However, the procedure is often incomplete owing to extensive vascular structure and arteriovenous shunts. We report our experience with embolization of these lesions by means of direct puncture and intratumoral injection of n-butyl cyanoacrylate (NBCA) or Onyx. Ten patients aged 32-82 years who were referred for preoperative embolization of seven carotid body tumors and three jugular paragangliomas were retrospectively analyzed. Intratumoral injections were primarily performed in four cases with multiple small-caliber arterial feeders and adjunctive to transarterial embolization in six cases with incomplete devascularization. Punctures were performed under ultrasound and injections were performed under roadmap fluoroscopic guidance. Detailed angiographies were performed before and after embolization procedures. Control angiograms showed complete or near-complete devascularization in all tumors. Three tumors with multiple small-caliber arterial feeders were treated with primary NBCA injections. One tumor necessitated transarterial embolization after primary injection of Onyx. Six tumors showed regional vascularization from the vasa vasorum or small-caliber branches of the external carotid artery following the transarterial approach. These regions were embolized with NBCA injections. No technical or clinical complications related to embolization procedures occurred. All except one of the tumors were surgically removed following embolization. In conclusion, preoperative devascularization with percutaneous direct injection of NBCA or Onyx is feasible, safe, and effective in head and neck paragangliomas with multiple small-caliber arterial feeders and in cases of incomplete devascularization following transarterial embolization.

  1. Silicon Promotes Adventitious Shoot Regeneration and Enhances Salinity Tolerance of Ajuga multiflora Bunge by Altering Activity of Antioxidant Enzyme

    PubMed Central

    Sivanesan, Iyyakkannu; Jeong, Byoung Ryong

    2014-01-01

    We investigated the effect of Si concentration on shoot regeneration and salinity tolerance of Ajuga multiflora. Addition of Si to the shoot induction medium significantly increased the frequency of shoot induction. The average number of shoots regenerated per explant decreased on the medium containing NaCl alone, while there was less decrease when the shoot induction medium was supplemented with both NaCl and Si. The shoot induction percentage increased linearly with increasing concentration of Si in the NaCl containing medium. Addition of Si to the shoot induction medium significantly increased SOD, POD, APX, and CAT activity in regenerated shoot buds as compared with the control. The inclusion of Si to the NaCl containing medium significantly increased the SOD activity in leaves and roots, while it decreased POD, APX, and CAT activity in both organs. Scanning electron microscopic analysis showed that there are no distinct differences in the structure of stomata between the control and Si-treated plants. However, NaCl treatment significantly affected the structure and number of stomata as compared to the control. Wavelength dispersive X-ray analysis confirmed the high Si deposition in trichomes of plants grown in the Si containing medium but not in plants grown in the medium without Si. PMID:24526904

  2. In vitro adventitious shoot regeneration via indirect organogenesis from inflorescence explants and peroxidase involvement in morphogenesis of Populus euphratica Olivier.

    PubMed

    Zhou, Yan; Gao, Ziyang; Gao, Shumin; Sun, Fangfang; Cheng, Pengjun; Li, Fenglan

    2012-12-01

    The inflorescences as explants for rapid propagation in vitro remained unknown in Populus euphratica Olivier. Here, we reported that multiple shoots were initiation from calli of both male and female inflorescences. The optimum medium for shoot induction from male inflorescences was lactose sulfite medium containing 1.0 mg L(-1) 6-benzylaminopurine (BA) and 0.5 mg L(-1) α-naphthalene acetic acid (NAA) or Murashige and Skoog (MS) medium containing 0.5 mg L(-1) BA and 0.2 mg L(-1) NAA. The optimum medium of shoot induction from female inflorescence calli was the MS medium containing 0.5 mg L(-1) BA and 0.2 mg L(-1) NAA. Rooting of regenerated shoots was obtained on 1/2 MS medium supplemented with 0.5∼1.0 mg L(-1) indole-3-butyric acid (IBA) and the highest frequency rooting was on medium containing 0.5 mg L(-1) IBA. No shoots were obtained on medium without BA and NAA. Peroxidase (POD) activity was measured by polyacrylamide gel electrophoresis during shoot induction and differentiation stages. The results showed that two bands of POD (2a and 2b) activity appeared lowest during the early 8 days at the dedifferentiation phase of leaves inducing calli, whereas POD 2a, 2b activity appeared to be increasing at the homeochronous dedifferentiation phase of inflorescence. Five most intensive bands, POD 1a, 1b, 1c, 2a, and ab, appeared in 8th and 28th days at the redifferentiation phase during shoot morphogenesis. These results demonstrated that the POD was involved in shoot morphogenesis from both leaf and inflorescence explants of Populus euphratica.

  3. [Cartography and geographical spread of the adventitious species of brome (Bromus spp.) among cereals in the Sais area of Morocco].

    PubMed

    Hamal, A; Benbella, M; Rzozi, S B; Bouhache, M; Msatef, Y

    2001-01-01

    Bromus spp is causing serious problems in wheat in the Sais area. However, the damage of this weed varies from one region to another according to the agro-climatic conditions and crop systems. The characterization of the infestation level in each situation is a prerequisite to develop a control strategy adapted to each environment. This study was undertaken in order to determine the infestation level and geographical spread of the weedy brome (Bromus spp) on wheat in Sais following crop systems and pedo-climatic conditions. The results obtained during two consecutive years (1998-99 and 1999-2000) revealed that ripgut brome (Bromus rigidus Roth.) was the most dominant species in wheat fields in the surveyed regions, followed by B. rubens L., B. sterilis L., B. madritensis L. and B. mollus L. Among, 18 regions and 100 infested wheat fields, 16.67% of fields were slightly infested (Plant density of Bromus (Dbr < 90 plants/m2, 61.11% were moderately infested (90 < Dbr < 290 plants/m2) and 22.22% were highly infested (Dbr > 400 plants/m2). The maximum relative frequency was obtained with Bromus rigidus (47.15%) and the coverage was 40.43%. But, for B.rubens, B. madritensis and B. sterilis, the relative frequencies were respectively 31.42; 26 and 15% and their coverages were respectively 28.9, 20.4 and 12.5%. PMID:12425101

  4. [Cartography and geographical spread of the adventitious species of brome (Bromus spp.) among cereals in the Sais area of Morocco].

    PubMed

    Hamal, A; Benbella, M; Rzozi, S B; Bouhache, M; Msatef, Y

    2001-01-01

    Bromus spp is causing serious problems in wheat in the Sais area. However, the damage of this weed varies from one region to another according to the agro-climatic conditions and crop systems. The characterization of the infestation level in each situation is a prerequisite to develop a control strategy adapted to each environment. This study was undertaken in order to determine the infestation level and geographical spread of the weedy brome (Bromus spp) on wheat in Sais following crop systems and pedo-climatic conditions. The results obtained during two consecutive years (1998-99 and 1999-2000) revealed that ripgut brome (Bromus rigidus Roth.) was the most dominant species in wheat fields in the surveyed regions, followed by B. rubens L., B. sterilis L., B. madritensis L. and B. mollus L. Among, 18 regions and 100 infested wheat fields, 16.67% of fields were slightly infested (Plant density of Bromus (Dbr < 90 plants/m2, 61.11% were moderately infested (90 < Dbr < 290 plants/m2) and 22.22% were highly infested (Dbr > 400 plants/m2). The maximum relative frequency was obtained with Bromus rigidus (47.15%) and the coverage was 40.43%. But, for B.rubens, B. madritensis and B. sterilis, the relative frequencies were respectively 31.42; 26 and 15% and their coverages were respectively 28.9, 20.4 and 12.5%.

  5. Vascular response to stress in health and disease.

    PubMed

    Plante, Gérard E

    2002-06-01

    The body's vasculature plays a critical role in the development of functional and structural alterations responsible for tissue and organ damage in laboratory animals and human subjects during illness and senescence, and in response to stress. Components of the vasculature, namely, major arteries such as the aorta, smaller arteries, arterioles, capillaries, post-capillary venules, and collecting central veins, all serve as conduits through which vital substrates are delivered to cellular masses and/or waste products are removed. A number of physical and neurohumoral agents known to be responsive to stress stimuli exert functional control over the vasculature. Both physical and emotional stress have been found to cause significant hemodynamic alterations. Large artery rigidity, for instance, develops rapidly following stress-induced activation of the autonomic nervous system. Associated with this process is increased release into the circulation of catecholamines and angiotensin-II. At the same time, insulin resistance develops, accompanied by nitric oxide release and changes in the immune system. The response of large arterial conduits to stress is characterized by increased pulse pressure, which in turn affects the endothelium of the arterial vessels responsible for determining total peripheral resistance. Microcirculation networks, where a large fraction of the blood volume is contained, are affected as well, and the blood in them is subject to redistribution into adjacent interstitial fluid compartments. Changes in endothelial permeability, secondary to variations in pulse pressure, can lead to interstitial edema and changes in the physicochemical properties of interstitial compartments. These changes give rise to alterations in the traffic of substrates and waste products between blood and cells. This sequence of events also takes place in the vasa vasorum microcirculation that nourishes large arteries, and likely contributes to remodeling of the vascular wall

  6. Adventitious Presence of Patented Genetically Modified Organisms on Private Premises: Is Intent Necessary for Actions in Infringement against the Property Owner?

    ERIC Educational Resources Information Center

    Mgbeoji, Ikechi

    2007-01-01

    The law of patents has long struggled with the status of intent in determining liability for infringement. This struggle has recently been given a sharper edge by the emergence of biotechnological products with the inherent ability of auto-dispersal and regeneration. The question thus is whether a person on whose backyard a patented genetic…

  7. No Identical "Mesenchymal Stem Cells" at Different Times and Sites: Human Committed Progenitors of Distinct Origin and Differentiation Potential Are Incorporated as Adventitial Cells in Microvessels.

    PubMed

    Sacchetti, Benedetto; Funari, Alessia; Remoli, Cristina; Giannicola, Giuseppe; Kogler, Gesine; Liedtke, Stefanie; Cossu, Giulio; Serafini, Marta; Sampaolesi, Maurilio; Tagliafico, Enrico; Tenedini, Elena; Saggio, Isabella; Robey, Pamela G; Riminucci, Mara; Bianco, Paolo

    2016-06-14

    A widely shared view reads that mesenchymal stem/stromal cells ("MSCs") are ubiquitous in human connective tissues, can be defined by a common in vitro phenotype, share a skeletogenic potential as assessed by in vitro differentiation assays, and coincide with ubiquitous pericytes. Using stringent in vivo differentiation assays and transcriptome analysis, we show that human cell populations from different anatomical sources, regarded as "MSCs" based on these criteria and assumptions, actually differ widely in their transcriptomic signature and in vivo differentiation potential. In contrast, they share the capacity to guide the assembly of functional microvessels in vivo, regardless of their anatomical source, or in situ identity as perivascular or circulating cells. This analysis reveals that muscle pericytes, which are not spontaneously osteochondrogenic as previously claimed, may indeed coincide with an ectopic perivascular subset of committed myogenic cells similar to satellite cells. Cord blood-derived stromal cells, on the other hand, display the unique capacity to form cartilage in vivo spontaneously, in addition to an assayable osteogenic capacity. These data suggest the need to revise current misconceptions on the origin and function of so-called "MSCs," with important applicative implications. The data also support the view that rather than a uniform class of "MSCs," different mesoderm derivatives include distinct classes of tissue-specific committed progenitors, possibly of different developmental origin. PMID:27304917

  8. In vitro antifungal activity of extracts obtained from Hypericum perforatum adventitious roots cultured in a mist bioreactor against planktonic cells and biofilm of Malassezia furfur.

    PubMed

    Simonetti, Giovanna; Tocci, Noemi; Valletta, Alessio; Brasili, Elisa; D'Auria, Felicia Diodata; Idoux, Alicia; Pasqua, Gabriella

    2016-01-01

    Xanthone-rich extracts from Hypericum perforatum root cultures grown in a Mist Bioreactor as antifungal agents against Malassezia furfur. Extracts of Hypericum perforatum roots grown in a bioreactor showed activity against planktonic cells and biofilm of Malassezia furfur. Dried biomass, obtained from roots grown under controlled conditions in a ROOTec mist bioreactor, has been extracted with solvents of increasing polarity (i.e. chloroform, ethyl acetate and methanol). The methanolic fraction was the richest in xanthones (2.86 ± 0.43 mg g(-1) DW) as revealed by HPLC. The minimal inhibitory concentration of the methanol extract against M. furfur planktonic cells was 16 μg mL(-1). The inhibition percentage of biofilm formation, at a concentration of 16 μg mL(-1), ranged from 14% to 39%. The results show that H. perforatum root extracts could be used as new antifungal agents in the treatment of Malassezia infections.

  9. Data on the parasitological status of golden jackal (Canis aureus L., 1758) in Hungary.

    PubMed

    Takács, András; Szabó, László; Juhász, Lajos; Takács, András Attila; Lanszki, József; Takács, Péter Tamás; Heltai, Miklós

    2014-03-01

    In Hungary, twenty Canis aureus individuals were submitted to parasitological examinations in 2010-2012. Two Coccidia: Cystoisospora canis (15%) and Toxoplasma-type oocysts (5%), one Trematoda: Alaria alata (10%), six Cestoda: Mesocestoides lineatus (20%), Echinococcus granulosus (10%), Dipylidium caninums (5%), Taenia hydatigena (15%), Taenia pisiformis (20%), Taenia crassiceps (40%), and nine Nematoda: Angiostrongylus vasorum (10%), Crenosoma vulpis (30%), Capillaria aerophila (5%), Toxocara canis (20%), Toxascaris leonina (15%), Trichuris vulpis (10%), Ancylostoma caninum (45%), Uncinaria stenocephala (40%), Capillaria plica (45%) have been identified. Angiostronglyus vasorum has been reported from carnivores in Europe, Africa, South America and North America. The helminth A. vasorum or French heartworm is a metastrongylid nematode, widely distributed in Western Europe, that infects the pulmonary arterial tree of dogs, various species of foxes, wolves, Eurasian badgers, coyotes and stoats. To our knowledge, this is the first report of natural A. vasorum infection in golden jackal. PMID:24334089

  10. New insights onto cardiopulmonary nematodes of dogs in Sardinia, Italy.

    PubMed

    Pipia, A P; Varcasia, A; Tosciri, G; Seu, S; Manunta, M L; Mura, M C; Sanna, G; Tamponi, C; Brianti, E; Scala, A

    2014-04-01

    Dog heartworms Angiostrongylus vasorum and Dirofilaria immitis cause severe parasitological diseases; the importance of these parasitosis is growing due to their health impact on animals, the possible zoonotic implications and the recent spreading across several European countries and previously non-endemic areas. The aim of this study is to update the epidemiological scenario of cardiopulmonary nematodes A. vasorum and D. immitis in dogs of Sardinia island and to perform a morphological identification of larvae by the use of the Baermann and Knott techniques respectively and the molecular characterization of the mitochondrial cytochrome c oxidase subunit I (cox1) and the second ribosomal transcribed spacer region (ITS-2) of larvae L1 of A. vasorum. In the present study, 3.4% (5/146) of dogs resulted positive at Baermann technique for A. vasorum while 8.9% (61/684) to D. immitis. If on one side A. vasorum can be considered an emerging parasite in Sardinia, the parasitic pressure and the risk of infection for D. immitis in the island seems to be increased compared with the recent past.

  11. Melatonin promotes adventitious root regeneration in in vitro shoot tip explants of the commercial sweet cherry rootstocks CAB-6P (Prunus cerasus L.), Gisela 6 (P. cerasus × P. canescens), and MxM 60 (P. avium × P. mahaleb).

    PubMed

    Sarropoulou, Virginia N; Therios, Ioannis N; Dimassi-Theriou, Kortessa N

    2012-01-01

    The objectives of this study were to test the effects of melatonin (N-acetyl-5-methoxytryptamine), a natural compound of edible plants on the rooting of certain commercial sweet cherry rootstocks. Shoot tip explants from previous in vitro cultures of the cherry rootstocks CAB-6P (Prunus cerasus L.), Gisela 6 (P. cerasus × P. canescens), and M × M 60 (P. avium × P. mahaleb) were included in the experiment. The effect of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) alone or in combination with melatonin was tested concerning their rooting potential. Seven concentrations of melatonin (0, 0.05, 0.1, 0.5, 1, 5, and 10 μM) alone or in combination with 5.71 μM of IAA or 4.92 μM of IBA were tested. For each rootstock, 21 treatments were included. The explants were grown in glass tubes containing 10 mL of substrate. The parameters measured include rooting percentage, number of roots per rooted explant, root length, and callus formation. The data presented in this study show that melatonin has a rooting promoting effect at a low concentration but a growth inhibitory effect at high concentrations. In the absence of auxin, 1 μM melatonin had auxinic response concerning the number and length of roots, but 10 μM melatonin was inhibitory to rooting in all the tested rootstocks. The final conclusion of this experiment is that exogenously applied melatonin acted as a rooting promoter and its action was similar to that of IAA. PMID:21749439

  12. Analysis of gene expression profiles for cell wall modifying proteins and ACC synthases in soybean cyst nematode colonized roots, adventitious rooting hypocotyls, root tips, flooded roots, and IBA and ACC treatment roots

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We hypothesized that soybean cyst nematode (SCN) co-opts a part or all of one or more innate developmental process in soybean to establish its feeding structure, syncytium, in soybean roots. The syncytium in soybean roots is formed in a predominantly lateral direction within the vascular bundle by ...

  13. Growth responses and adaptations of Fraxinus pennsylvanica seedlings to flooding

    SciTech Connect

    Sena Gomes, A.R.; Kozlowski, T.T.

    1980-01-01

    Flooding induced several physiological and morphological changes in Fraxinus pennsylvanica seedlings, with stomatal closure among the earliest responses. Subsequent changes included: reduction in dry weight increment of roots, stems, and leaves; formation of hypertrophied lenticles and production of adventitious roots on submerged portions of the stem above the soil line; leaf necrosis; and leaf abscission. After 15 days of stomatal closure as a results of flooding, stomata began to reopen progressively until stomata aperture was similar in flooded and unflooded plants. Adventitious roots began to form at about the time stomatal reopening began. As more adventitious roots formed, elongated, and branched, the stomata opened further. The formation of adventitious roots was in important adaptation for flooding tolerance as shown by the high efficiency of adventitious roots in absorption of water and in high correlation between the production of adventitious roots and stomatal reopening. 6 figures, 2 tables.

  14. UAP56 Couples piRNA Clusters to the Perinuclear Transposon Silencing Machinery

    PubMed Central

    Zhang, Fan; Wang, Jie; Xu, Jia; Zhang, Zhao; Koppetsch, Birgit S.; Schultz, Nadine; Vreven, Thom; Meignin, Carine; Davis, Ilan; Zamore, Phillip D.; Weng, Zhiping; Theurkauf, William E.

    2012-01-01

    Summary piRNAs silence transposons during germline development. In Drosophila, transcripts from heterochromatic clusters are processed into primary piRNAs in the perinuclear nuage. The nuclear DEAD box protein UAP56 has been previously implicated in mRNA splicing and export, while the DEAD box protein Vasa has an established role in piRNA production and localizes to nuage with the piRNA binding PIWI proteins Ago3 and Aub. We show that UAP56 co-localizes with the cluster-associated HP1 variant Rhino, that nuage granules containing Vasa localize directly across the nuclear envelope from cluster foci containing UAP56 and Rhino, and that cluster transcripts immunoprecipitate with both Vasa and UAP56. Significantly, a charge-substitution mutation that alters a conserved surface residue in UAP56 disrupts co-localization with Rhino, germline piRNA production, transposon silencing, and perinuclear localization of Vasa. We therefore propose that UAP56 and Vasa function in a piRNA-processing compartment that spans the nuclear envelope. PMID:23141543

  15. Changes within vascular bundles of rodent kidneys caused by different diets.

    PubMed

    Yaakobi, D; Borut, A

    1977-07-11

    Morphological effects caused by two different diets (low protein-high water intake, and high protein-restricted water) on the vascular bundles in the outer medullary zone of the kidney were studied in the laboratory white mouse and in the golden spiny mouse (Acomys russatus, Muridae). In both rodents, when on a low protein-high water intake diet, considerable interstitial substance was found between the vasa recta of the bundles. No interstitial substance was found in animals on high protein-low water intake diet; as a result the vasa recta of the vascular bundle adhered closely. The low protein-high water intake diet caused a marked diuresis, low urine osmolality and low urinary urea concentration. It is assumed that the increase in interstitial substance between the vasa recta of the vascular bundle lowers the efficiency of the counter current barrier system for urea in the kidney and, as a consequence, the medullary urea gradient and urine concentrating capacity decreases. In animals on a high protein diet, the closely juxtaposed vasa recta assure an efficient countercurrent exchange, leading to accumulation and maintenance of a large urea gradient in the medulla and maximal urine concentration. It is suggested that the amount of interstitial material between the vasa recta of the vascular bundle might serve as a modulating factor for the urea concentration in the kidney.

  16. Root formation in ethylene-insensitive plants.

    PubMed

    Clark, D G; Gubrium, E K; Barrett, J E; Nell, T A; Klee, H J

    1999-09-01

    Experiments with ethylene-insensitive tomato (Lycopersicon esculentum) and petunia (Petunia x hybrida) plants were conducted to determine if normal or adventitious root formation is affected by ethylene insensitivity. Ethylene-insensitive Never ripe (NR) tomato plants produced more below-ground root mass but fewer above-ground adventitious roots than wild-type Pearson plants. Applied auxin (indole-3-butyric acid) increased adventitious root formation on vegetative stem cuttings of wild-type plants but had little or no effect on rooting of NR plants. Reduced adventitious root formation was also observed in ethylene-insensitive transgenic petunia plants. Applied 1-aminocyclopropane-1-carboxylic acid increased adventitious root formation on vegetative stem cuttings from NR and wild-type plants, but NR cuttings produced fewer adventitious roots than wild-type cuttings. These data suggest that the promotive effect of auxin on adventitious rooting is influenced by ethylene responsiveness. Seedling root growth of tomato in response to mechanical impedance was also influenced by ethylene sensitivity. Ninety-six percent of wild-type seedlings germinated and grown on sand for 7 d grew normal roots into the medium, whereas 47% of NR seedlings displayed elongated tap-roots, shortened hypocotyls, and did not penetrate the medium. These data indicate that ethylene has a critical role in various responses of roots to environmental stimuli.

  17. Ischemic Effects of Transcatheter Arterial Embolization with N-Butyl Cyanoacrylate-Lipiodol on the Colon in a Swine Model

    SciTech Connect

    Ikoma, Akira; Kawai, Nobuyuki; Sato, Morio Sonomura, Tetsuo; Minamiguchi, Hiroki; Nakai, Motoki; Takasaka, Isao; Nakata, Kouhei; Sahara, Shinya; Sawa, Naohisa; Shirai, Shintaro; Mori, Ichiro

    2010-10-15

    This study was designed to assess the safety of transcatheter arterial embolization (TAE) with n-butyl cyanoacrylate-lipiodol (NBCA-Lp) for the large bowel and to investigate the vital response to NBCA-Lp in a swine model. In nine swine, nine arteries nourishing the colon were embolized with NBCA-Lp (1 ml of NBCA mixed with 4 ml of lipiodol): sigmoid-rectal branch artery in six swine, right colic branch artery in two, and middle colic branch artery in one. The amount of NBCA-Lp was 0.1-0.4 ml. Sacrifice was conducted 3 days after TAE to identify histological infarction. Classification was conducted retrospectively: group A, vasa recta without NBCA-Lp embolization despite TAE; group B, three or fewer vasa recta with NBCA-Lp embolization; and group C, five or more vasa recta with NBCA-Lp embolization. In one swine in group A, no necrotic focus was observed. In group B, three of four swine experienced no ischemic damage. The remaining one swine experienced necrosis of mucosal and submucosal layers in one-fourth of the circumference. In group C, all four swine with marginal artery and five vasa recta or more embolized experienced total necrosis of mucosa, submucosa, and smooth muscle layers of the whole colonic circumference. Significant difference on the extent of ischemic damage was observed between groups B and C (P < 0.05). Microscopically, NBCA-Lp induced acute vasculitis. Embolization of three or fewer vasa recta with NBCA-Lp induced no ischemic damage or limited necrosis, whereas embolization of five or more vasa recta with NBCA-Lp induced extensive necrosis.

  18. The male reproductive system of Hippolyte inermis Leach 1815 (Decapoda, Caridea)

    NASA Astrophysics Data System (ADS)

    Cobos, Vanesa; Díaz, Vanessa; Raso, Jose Enrique García; Manjón-Cabeza, M. E.

    2011-03-01

    The present work completes a series of studies on the biology of the shrimp Hippolyte inermis Leach 1815, where we suggested the species to be gonochoristic. The morphology of the male reproductive system (testes, vasa deferentia, gonopores) and the different stages of male germ cell development are described for the first time in the genus Hippolyte, using TEM, SEM, and histological methods. All males from 1.70 to 3.42 mm in carapace length had active testes and well-developed vasa deferentia. No case of sex reversal could be found.

  19. Granulosa cells and retinoic acid co-treatment enrich potential germ cells from manually selected Oct4-EGFP expressing human embryonic stem cells.

    PubMed

    Chen, Hsin-Fu; Jan, Pey-Shynan; Kuo, Hung-Chih; Wu, Fang-Chun; Lan, Chen-Wei; Huang, Mei-Chi; Chien, Chung-Liang; Ho, Hong-Nerng

    2014-09-01

    Differentiation of human embryonic stem (HES) cells to germ cells may become clinically useful in overcoming diseases related to germ-cell development. Niches were used to differentiate HES cell lines, NTU1 and H9 Oct4-enhanced green fluorescence protein (EGFP), including laminin, granulosa cell co-culture or conditioned medium, ovarian stromal cell co-culture or conditioned medium, retinoic acid, stem cell factor (SCF) and BMP4-BMP7-BMP8b treatment. Flow cytometry showed that granulosa cell co-culture (P < 0.001) or conditioned medium (P = 0.007) treatment for 14 days significantly increased the percentages of differentiated H9 Oct4-EGFP cells expressing early germ cell marker stage-specific embryonic antigen 1(SSEA1); sorted SSEA1[+] cells did not express higher levels of germ cell gene VASA and GDF9. Manually collected H9 Oct4-EGFP[+] cells expressed significantly higher levels of VASA (P = 0.005) and GDF9 (P = 0.001). H9 Oct4-EGFP[+] cells developed to ovarian follicle-like structures after culture for 28 days but with low efficiency. Unlike SCF and BMP4, retinoic acid co-treatment enhanced VASA, GDF9 and SCP3 expression. A protocol is recommended to enrich differentiated HES cells with germ-cell potential by culture with granulosa cells, conditioned medium or retinoic acid, manual selection of Oct4-EGFP[+] cells, and analysis of VASA, GDF9 expression, or both.

  20. Reproductive morphology of Ornithonyssus sylviarum (Canestrini and Fanzago) (Acari: Macronyssidae).

    PubMed

    Pound, J M; Oliver, J H

    1976-06-01

    The male reproductive system consists of paired tests, vasa deferentia and dark-staining lateral accessory glands, single ventromedial spongy accessory gland, seminal vesicle, ejaculatory apparatus, and gonopore. The female system consists of paired rami sacculi, sacculus foemineus, ovary (lyriform and medial portions), oviduct, vagina, and paired vaginal glands. PMID:932922

  1. Improved production of ginsenosides in suspension cultures of ginseng by medium replenishment strategy.

    PubMed

    Jeong, Cheol-Seung; Murthy, Hosakatte Niranjana; Hahn, Eun-Joo; Paek, Kee-Yoeup

    2008-03-01

    The objective of this study was to improve the accumulation of ginsenosides by the adventitious root cultures of ginseng, which are important secondary metabolites with pharmaceutical applications. The adventitious roots were cultured in bioreactors for 50 d using 1.5-strength Murashige and Skoog (MS) medium supplemented with 10 mg/l indole acetic acid and 75 g/l sucrose. Kinetic studies of the nutrient composition of the spent medium revealed the gradual depletion of various inorganic nutrients and sugars. Cultures were supplied with fresh nutrient medium (medium exchange or replenishment with 0.75- and 1.0-strength MS medium) after 10 and 20 d of culture initiation to fulfill the nutritional requirements of adventitious roots. Medium replenishment strategy (with 1.0-strength MS medium after 20 d of culture) significantly improved the growth of adventitious roots and the biosynthesis of ginsenosides by the adventitious roots. This work is useful for the large-scale cultivation of adventitious roots for the production of ginsenosides. PMID:18397781

  2. Diversity and prevalence of metastrongyloid nematodes infecting the red panda (Ailurus fulgens) in European zoos.

    PubMed

    Bertelsen, Mads F; Meyland-Smith, Frederik; Willesen, Jakob L; Jefferies, Ryan; Morgan, Eric R; Monrad, Jesper

    2010-09-20

    Metastrongyloid induced pneumonia has been described sporadically in the red panda (Ailurus fulgens). Early descriptions in pandas recently imported to the USA from China involved parasites morphologically similar to Angiostrongylus spp. and Crenosomatidae. More recently, four cases of severe verminous pneumonia associated with Angiostrongylus vasorum have been reported from European zoos. A coprological survey of the red panda population within European zoos was conducted in 2008. Faecal samples from 115 pandas originating from 54 zoos were collected on 3 consecutive days. Using Baermann technique, 40 animals (35%) from 20 zoos (37%) were found to shed metastrongyloid first stage larvae (L(1)). Based on their morphology and size, the L(1) observed could be divided into three morphologically distinct types: (1) a Crenosoma sp. type (n=5, overall prevalence: 4.3%), (2) an A. vasorum type (n=3, 2.6%), and (3) an unidentified metastrongyloid species, similar to, but morphologically distinct from A. vasorum (n=32, 27.8%). Further confirmation of species identification was provided by PCR amplification and sequencing of the 18S rRNA gene, which confirmed three different species. The novel Crenosoma species was most genetically analogous to Crenosoma mephitidis and the unidentified metastrongyloid species was most similar to Stenurus minor and Torynurus convulutus. Routine and quarantine health care of red pandas in captivity should take account of the risk of Angiostrongylus and Crenosoma infection in endemic areas, but should also be cognisant of the widespread presence of an apparently less pathogenic species of lungworm. The identity of the two potentially novel species is subject to further work.

  3. Lung and hearth nematodes in some Spanish mammals.

    PubMed

    Alvarez, F; Iglesias, R; Bos, J; Rey, J; Sanmartin Durán, M L

    1991-01-01

    Thirteen host species belonging to the orders Rodentia, Insectivora and Carnivora from various localities in Galicia (NW Spain) were examined for heart and lung parasites. The following species were found: Parastrongylus dujardini (5.5%) in Apodemus sylvaticus, Crenosoma striatum in Erinaceus europaeus (83%), Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus in Vulpes vulpes (3, 3.46 and 0.50%, respectively), Crenosoma taiga in Putorius putorius (100%) and Crenosoma sp. in Meles meles (25%). In Crocidura russula nematode larvae were found (3.3%). Mus musculus, Rattus norvegicus, Rattus rattus, Talpa caeca, Sorex araneus, Genetta genetta and Canis lupus were not parasitized by lung or heart parasites.

  4. Regulation of organogenesis and somatic embryogenesis by auxin in melon, Cucumis melo L.

    PubMed

    Tabei, Y; Kanno, T; Nishio, T

    1991-08-01

    Various tissues of seeds and seedlings of melon were cultured in vitro to study the effects of auxin concentration on organogenesis and embryogenesis. Adventitious shoots and somatic embryos were formed from explants of cotyledons of mature seeds, hypocotyls of seedlings, and leaves and petioles of young plantlets. Expanded cotyledons of seedlings formed only adventitious shoots. All tissues responded similarly to the 2,4-D concentration in the media, that is, adventitious shoots were formed at low concentration, callus proliferated without differentiation at intermediate concentration and somatic embryos were induced at high concentration. Cotyledons of mature seeds formed both adventitious shoots and somatic embryos more efficiently than any other tissues cultured.Effects of three auxins, 2,4-D, NAA and IAA, on organogenesis and embryogenesis were compared using cotyledons of mature seeds. Adventitious shoots were formed at low level of auxins (0 to 0.01 mg/l 2,4-D; 0 to 0.1 mg/l NAA; 0 to 1.0 mg/l IAA), and embryos were formed at high level of auxins (1.0 to 2.0 mg/l 2,4-D; 3.0 to 10.0 mg/l NAA; 20.0 to 100.0 mg/l IAA). IAA gave more efficient shoot formation and embryogenesis than the other auxins. PMID:24221584

  5. A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering.

    PubMed

    Sebo, Zachary L; Lee, Han B; Peng, Ying; Guo, Yi

    2014-01-01

    The type II CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeats/CRISPR-associated) has recently emerged as an efficient and simple tool for site-specific engineering of eukaryotic genomes. To improve its applications in Drosophila genome engineering, we simplified the standard two-component CRISPR/Cas9 system by generating a stable transgenic fly line expressing the Cas9 endonuclease in the germline (Vasa-Cas9 line). By injecting vectors expressing engineered target-specific guide RNAs into Vasa-Cas9 fly embryos, mutations were generated from site-specific DNA cleavages and efficiently transmitted into progenies. Because Cas9 endonuclease is the universal component of the type II CRISPR/Cas9 system, site-specific genomic engineering based on this improved platform can be achieved with lower complexity and toxicity, greater consistency, and excellent versatility.

  6. The presence of Bt-transgenic oilseed rape in wild mustard populations affects plant growth.

    PubMed

    Liu, Yongbo; Stewart, C Neal; Li, Junsheng; Huang, Hai; Zhang, Xitao

    2015-12-01

    The adventitious presence of transgenic plants in wild plant populations is of ecological and regulatory concern, but the consequences of adventitious presence are not well understood. Here, we introduced Bacillus thuringiensis Cry1Ac (Bt)-transgenic oilseed rape (Bt OSR, Brassica napus) with various frequencies into wild mustard (Brassica juncea) populations. We sought to better understand the adventitious presence of this transgenic insecticidal crop in a wild-relative plant population. We assessed the factors of competition, resource availability and diamondback moth (Plutella xylostella) infestation on plant population dynamics. As expected, Bt OSR performed better than wild mustard in mixed populations under herbivore attack in habitats with enough resources, whereas wild mustard had higher fitness when Bt OSR was rarer in habitats with limited resources. Results suggest that the presence of insect-resistant transgenic plants could decrease the growth of wild mustard and Bt OSR plants and their populations, especially under high herbivore pressure.

  7. Branching out in roots: uncovering form, function, and regulation.

    PubMed

    Atkinson, Jonathan A; Rasmussen, Amanda; Traini, Richard; Voß, Ute; Sturrock, Craig; Mooney, Sacha J; Wells, Darren M; Bennett, Malcolm J

    2014-10-01

    Root branching is critical for plants to secure anchorage and ensure the supply of water, minerals, and nutrients. To date, research on root branching has focused on lateral root development in young seedlings. However, many other programs of postembryonic root organogenesis exist in angiosperms. In cereal crops, the majority of the mature root system is composed of several classes of adventitious roots that include crown roots and brace roots. In this Update, we initially describe the diversity of postembryonic root forms. Next, we review recent advances in our understanding of the genes, signals, and mechanisms regulating lateral root and adventitious root branching in the plant models Arabidopsis (Arabidopsis thaliana), maize (Zea mays), and rice (Oryza sativa). While many common signals, regulatory components, and mechanisms have been identified that control the initiation, morphogenesis, and emergence of new lateral and adventitious root organs, much more remains to be done. We conclude by discussing the challenges and opportunities facing root branching research. PMID:25136060

  8. The presence of Bt-transgenic oilseed rape in wild mustard populations affects plant growth.

    PubMed

    Liu, Yongbo; Stewart, C Neal; Li, Junsheng; Huang, Hai; Zhang, Xitao

    2015-12-01

    The adventitious presence of transgenic plants in wild plant populations is of ecological and regulatory concern, but the consequences of adventitious presence are not well understood. Here, we introduced Bacillus thuringiensis Cry1Ac (Bt)-transgenic oilseed rape (Bt OSR, Brassica napus) with various frequencies into wild mustard (Brassica juncea) populations. We sought to better understand the adventitious presence of this transgenic insecticidal crop in a wild-relative plant population. We assessed the factors of competition, resource availability and diamondback moth (Plutella xylostella) infestation on plant population dynamics. As expected, Bt OSR performed better than wild mustard in mixed populations under herbivore attack in habitats with enough resources, whereas wild mustard had higher fitness when Bt OSR was rarer in habitats with limited resources. Results suggest that the presence of insect-resistant transgenic plants could decrease the growth of wild mustard and Bt OSR plants and their populations, especially under high herbivore pressure. PMID:26338267

  9. Branching Out in Roots: Uncovering Form, Function, and Regulation1

    PubMed Central

    Atkinson, Jonathan A.; Rasmussen, Amanda; Traini, Richard; Voß, Ute; Sturrock, Craig; Mooney, Sacha J.; Wells, Darren M.; Bennett, Malcolm J.

    2014-01-01

    Root branching is critical for plants to secure anchorage and ensure the supply of water, minerals, and nutrients. To date, research on root branching has focused on lateral root development in young seedlings. However, many other programs of postembryonic root organogenesis exist in angiosperms. In cereal crops, the majority of the mature root system is composed of several classes of adventitious roots that include crown roots and brace roots. In this Update, we initially describe the diversity of postembryonic root forms. Next, we review recent advances in our understanding of the genes, signals, and mechanisms regulating lateral root and adventitious root branching in the plant models Arabidopsis (Arabidopsis thaliana), maize (Zea mays), and rice (Oryza sativa). While many common signals, regulatory components, and mechanisms have been identified that control the initiation, morphogenesis, and emergence of new lateral and adventitious root organs, much more remains to be done. We conclude by discussing the challenges and opportunities facing root branching research. PMID:25136060

  10. Two SCARECROW-LIKE genes are induced in response to exogenous auxin in rooting-competent cuttings of distantly related forest species.

    PubMed

    Sánchez, Conchi; Vielba, Jesús M; Ferro, Enrique; Covelo, Guillermo; Solé, Alicia; Abarca, Dolores; de Mier, Belén S; Díaz-Sala, Carmen

    2007-10-01

    We characterized SCARECROW-LIKE genes induced by auxin in rooting-competent cuttings of two distantly related forest species (Pinus radiata D. Don and Castanea sativa Mill.) before the activation of cell division that results in adventitious root formation. The predicted protein sequences contain domains characteristic of the GRAS protein family and show a strong similarity to the SCARECROW-LIKE proteins, indicating conserved functions of these proteins. Quantitative RT-PCR analysis showed that these genes are expressed at relatively high levels in roots. Induction of increased mRNA levels in rooting-competent cuttings of both species in response to exogenous auxin was observed within the first 24 h of the root induction process, a time when cell reorganization takes place, but before the resumption of cell division and the appearance of adventitious root primordia. These results suggest that SCARECROW-LIKE genes play a role during the earliest stages of adventitious root formation.

  11. Extraintestinal nematodes of the red fox Vulpes vulpes in north-west Italy.

    PubMed

    Magi, M; Guardone, L; Prati, M C; Mignone, W; Macchioni, F

    2015-07-01

    Extraintestinal nematodes of the red fox (Vulpes vulpes) are a wide group of parasites that infect wild and domestic carnivores and occasionally humans. Nematodes in the cardiopulmonary system, stomach, urinary apparatus and muscle tissue of 165 red foxes (Vulpes vulpes) from north-west Italy (Liguria and Piedmont) were investigated between 2009 and 2012. Of the cardiopulmonary nematodes, a high prevalence of Angiostrongylus vasorum and Eucoleus aerophilus (syn. Capillaria aerophila) was found, 78.2% and 41.8% respectively; Crenosoma vulpis (15.8%) and Filaroides spp. (4.8%) were also found. Spirocerca lupi (23.5%), Aonchotheca putorii (syn. Capillaria putorii) (8.6%) and Physaloptera spp. (2.5%) were detected in the stomach and Pearsonema plica (syn. Capillaria plica) (56.8%) in the bladder. Eucoleus boehmi (syn. Capillaria boehmi) was also detected in the nasal cavities of one of the two foxes examined. A coprological examination revealed eggs of E. aerophilus, A. putorii, S. lupi, Physaloptera spp. and eggs of intestinal parasites. Filarial worms were absent in all the 165 animals examined, nor was there evidence of Trichinella spp. in any of the foxes. The foxes were found to host a high prevalence of many species of extraintestinal nematodes. The prevalence of A. vasorum in foxes found in the present study is among the highest in Europe. In addition, to the best of our knowledge, E. boehmi and Filaroides spp. have never been reported before in this host in Italy.

  12. Molecular phylogeography of Angiostrongylus cantonensis (Nematoda: Angiostrongylidae) and genetic relationships with congeners using cytochrome b gene marker.

    PubMed

    Yong, Hoi-Sen; Eamsobhana, Praphathip; Song, Sze-Looi; Prasartvit, Anchana; Lim, Phaik-Eem

    2015-08-01

    Angiostrongylus cantonensis is an important emerging zoonotic parasite causing human eosinophilic meningitis (or meningoencephalitis) in many parts of the world. To-date there is only a single study using mitochondrial cytochrome b (CYTB) gene to determine its genetic structure in eight geographical localities in Thailand. The present study examined the molecular phylogeography of this rat lungworm and its phylogenetic relationship with congeners using CYTB gene marker. A total of 15 CYTB haplotypes was found in 37 sequences from 14 geographical localities (covering north, west, east, central and south regions) in Thailand. These CYTB haplotypes were distinct from those of A. cantonensis for China and Hawaii. In Thailand, some CYTB haplotypes appeared to be confined to specific geographical localities. The partial CYTB DNA nucleotide sequences separated unequivocally the A. cantonensis isolates of Thailand, China and Hawaii as well as the congeners Angiostrongylus malaysiensis, A. costaricensis and Angiostrongylus vasorum, with A. malaysiensis grouped with A. cantonensis and A. costaricensis grouped with A. vasorum. Likewise the congeners of Metastrongylus and Onchocerca genera could also be clearly differentiated. The present study added two new definitive hosts (Bandicota savilei and Rattus losea) and three new localities (Mae Hong Son in the north, Tak in the west, and Phang Nga in the south) for A. malaysiensis in Thailand, indicating its wide occurrence in the country. Three CYTB haplotypes were found in the Thailand samples of A. malaysiensis. In addition to differentiation of congeners, CYTB gene marker could be used for determining the genetic diversity of a given population/taxon.

  13. Development of interspecies testicular germ-cell transplantation in flatfish.

    PubMed

    Pacchiarini, Tiziana; Sarasquete, Carmen; Cabrita, Elsa

    2014-06-01

    Interspecific testicular germ cell (TGC) transplantation was investigated in two commercial flatfish species. Testes from donor species (Senegalese sole) were evaluated using classical histological techniques (haematoxylin-eosin staining and haematoxylin-light green-orange G-acid fuchsine staining), in situ hybridisation and immunohistochemical analysis. Both Ssvasa1-2 mRNAs and SsVasa protein allowed the characterisation of TGCs, confirming the usefulness of the vasa gene in the detection of Senegalese sole TGCs. Xenogenic transplants were carried out using TGCs from one-year-old Senegalese sole into turbot larvae. Propidium iodide-SYBR-14 and 4',6'-diamidino-2-phenylindole (DAPI) staining showed that 87.98% of the extracted testicular cells were viable for microinjection and that 15.63% of the total recovered cells were spermatogonia. The vasa gene was characterised in turbot recipients using cDNA cloning. Smvasa mRNA was confirmed as a germ cell-specific molecular marker in this species. Smvasa expression analysis during turbot ontogeny was carried out before Senegalese sole TGC transplants into turbot larvae. Turbot larvae at 18 days after hatching (DAH) proved to be susceptible to manipulation procedures. High survival rates (83.75±15.90-100%) were obtained for turbot larvae at 27, 34 and 42 DAH. These data highlight the huge potential of this species for transplantation studies. Quantitative PCR was employed to detect Senegalese sole vasa mRNAs (Ssvasa1-2) in the recipient turbot larvae. The Ssvasa mRNAs showed a significant increase in relative expression in 42-DAH microinjected larvae three weeks after treatment, showing the proliferation of Senegalese sole spermatogonia in transplanted turbot larvae.

  14. Fishery induces sperm depletion and reduction in male reproductive potential for crab species under male-biased harvest strategy.

    PubMed

    Pardo, Luis Miguel; Rosas, Yenifer; Fuentes, Juan Pablo; Riveros, Marcela Paz; Chaparro, Oscar Roberto

    2015-01-01

    Sperm depletion in males can occur when polygynous species are intensively exploited under a male-biased management strategy. In fisheries involving crabs species, the effects of this type of management on the reproductive potential is far from being understood. This study tests whether male-biased management of the principal Chilean crab fishery is able to affect the potential capacity of Metacarcinus edwardsii males to transfer sperm to females. Five localities in southern Chile, recording contrasting crab fishery landing, were selected to assess the potential of sperm depletion triggered by fishery. Seasonally, male crabs from each locality were obtained. Dry weight and histological condition of vasa deferentia and the Vaso-Somatic Index (VSI) were determined in order to use them as proxies for sperm depletion and male reproductive condition. A manipulative experiment was performed in the laboratory to estimate vasa deferentia weight and VSI from just-mated males in order to obtain a reference point for the potential effects of the fishery on sperm reserves. Sperm storage capacity is significantly affected by fisheries; during the mating season vasa deferentia from localities with low fishery intensity were heavier than those from high intensity fisheries, and these differences were even more evident in large males. Histological section showed that this disparity in vasa deferentia weight was explained principally by differences in the quantity of spermatophores rather than other seminal material. VSI was always higher in males from localities with low fishery intensity. Males from localities with high fishery intensity showed little capacity to recover sperm reserves and the VSI of these males remained below the values of the just-mated males. Detriment in the capacity of males to transfer sperm is the first step to sperm limitation in an exploited population, thus detection of sperm depletion can be an alert to introduce changes in the current management of

  15. Fishery Induces Sperm Depletion and Reduction in Male Reproductive Potential for Crab Species under Male-Biased Harvest Strategy

    PubMed Central

    Pardo, Luis Miguel; Rosas, Yenifer; Fuentes, Juan Pablo; Riveros, Marcela Paz; Chaparro, Oscar Roberto

    2015-01-01

    Sperm depletion in males can occur when polygynous species are intensively exploited under a male-biased management strategy. In fisheries involving crabs species, the effects of this type of management on the reproductive potential is far from being understood. This study tests whether male-biased management of the principal Chilean crab fishery is able to affect the potential capacity of Metacarcinus edwardsii males to transfer sperm to females. Five localities in southern Chile, recording contrasting crab fishery landing, were selected to assess the potential of sperm depletion triggered by fishery. Seasonally, male crabs from each locality were obtained. Dry weight and histological condition of vasa deferentia and the Vaso-Somatic Index (VSI) were determined in order to use them as proxies for sperm depletion and male reproductive condition. A manipulative experiment was performed in the laboratory to estimate vasa deferentia weight and VSI from just-mated males in order to obtain a reference point for the potential effects of the fishery on sperm reserves. Sperm storage capacity is significantly affected by fisheries; during the mating season vasa deferentia from localities with low fishery intensity were heavier than those from high intensity fisheries, and these differences were even more evident in large males. Histological section showed that this disparity in vasa deferentia weight was explained principally by differences in the quantity of spermatophores rather than other seminal material. VSI was always higher in males from localities with low fishery intensity. Males from localities with high fishery intensity showed little capacity to recover sperm reserves and the VSI of these males remained below the values of the just-mated males. Detriment in the capacity of males to transfer sperm is the first step to sperm limitation in an exploited population, thus detection of sperm depletion can be an alert to introduce changes in the current management of

  16. The pathogenesis of non-freezing cold nerve injury. Observations in the rat.

    PubMed

    Jia, J; Pollock, M

    1997-04-01

    Non-freezing cold nerve injury is uncommon in civilian practice, but may reach epidemic proportions in war zones. Studied since the time of Hippocrates, its aetiology has remained elusive. We sought to replicate experimentally, a peripheral nerve cold temperature gradient, since this has been emphasized in clinical descriptions. Our observations, in the rat, of the vasa nervorum show that cold-induced intravascular aggregation is followed by a 'no-reflow' phenomenon which culminates in endothelial damage and delayed thrombotic occlusion.

  17. The energy requirements for the basal efflux of 3H-noradrenaline from sympathetically innervated organs.

    PubMed

    Russ, H; Schömig, E; Trendelenburg, U

    1991-09-01

    In the rat vas deferens (preloaded with 3H-noradrenaline, catechol-O-methyl transferase inhibited, calcium-free solution) ouabain, glucose deprivation or the combination of hypoxia plus presence of lactate were found to induce a carrier-mediated (desipramine-sensitive) outward transport of the 3H-amine. Glucose deprivation additionally increased the efflux of deaminated 3H-metabolites, as a consequence of an increased net leakage of vesicular 3H-noradrenaline; moreover, 3H-dihydroxymandelic acid then became the predominant neuronal metabolite. The simultaneous lack of oxygen and glucose resulted in a very pronounced release of the 3H-amine. Moreover, during spontaneous efflux more outward transport of 3H-noradrenaline was observed in the absence than in the presence of extracellular calcium. In rat atria (under the same experimental conditions) the contribution by carrier-mediated outward transport to the spontaneous efflux of tritium exceeded that in vasa deferentia. Moreover, the efflux of lactate (as an index of hypoxia of the tissue) exceeded that observed in vasa deferentia, under aerobic and anaerobic conditions. It is proposed that the greater contribution by outward transport of 3H-noradrenaline to spontaneous efflux in atria than in vasa deferentia does not reflect any basic difference between the varicosities in two different organs. It is likely that the less heterogeneous distribution of the 3H-amine in atria than in vasa deferentia is responsible for storage of the exogenous amine in atrial varicosities that are subject to some hypoxia, to an increased extracellular lactate level and to perhaps a minor degree of glucose deficiency; these factors may well be responsible for the difference with regard to outward transport of 3H-noradrenaline during spontaneous efflux. Thus, in addition to the heterogeneity of the distribution of 3H-noradrenaline, an additional heterogeneity with regard to the energy supply must be expected for incubated organs.

  18. Verbal/Social Autopsy in Niger 2012–2013: A new tool for a better understanding of the neonatal and child mortality situation

    PubMed Central

    Bensaïd, Khaled; Yaroh, Asma Gali; Kalter, Henry D; Koffi, Alain K; Amouzou, Agbessi; Maina, Abdou; Kazmi, Narjis

    2016-01-01

    Niger, one of the poorest countries in the world, recently used for the first time the integrated verbal and social autopsy (VASA) tool to assess the biological causes and social and health system determinants of neonatal and child deaths. These notes summarize the Nigerien experience in the use of this new tool, the steps taken for high level engagement of the Niger government and stakeholders for the wide dissemination of the study results and their use to support policy development and maternal, neonatal and child health programming in the country. The experience in Niger reflects lessons learned by other developing countries in strengthening the use of data for evidence–based decision making, and highlights the need for the global health community to provide continued support to country data initiatives, including the collection, analysis, interpretation and utilization of high quality data for the development of targeted, highly effective interventions. In Niger, this is supporting the country’s progress toward achieving Millennium Development Goal 4. A follow–up VASA study is being planned and the tool is being integrated into the National Health Management Information System. VASA studies have now been completed or are under way in additional sub–Saharan African countries, in each through the same collaborative process used in Niger to bring together health policy makers, program planners and development partners. PMID:26955472

  19. The effect of urea infusion on the urinary concentrating mechanism in protein-depleted rats.

    PubMed Central

    Pennell, J P; Sanjana, V; Frey, N R; Jamison, R L

    1975-01-01

    To explore the role of urea in the urinary concentrating mechanism, the contents of vasa recta, Henle's descending limbs and collecting ducts were sampled by micropuncture of the renal papilla before and after infusion of urea in 10 protein-depleted rats. Eight protein-depleted rats not given urea were similarly studied as a control group. After urea administration, osmolality and the concentrations of urea and nonurea solute of urine from both exposed and contralateral kideny increased significantly. The osmolality and urea concentration of fluid from the end of Henle's descending limb and vasa recta plasma and the tubule fluid-to-plasma inulin ratio in the end-descending limb all increased significantly after urea infusion. We interpret these observations to indicate that urea enhances urinary concentration by increasing the abstraction of water from the juxtamedullary nephron (presumably the descending limb), in agreement with the prediction of recent passive models of the urinary concentrating mechanism. However, the concentration of urea in fluid from the descending limb after urea infusion was high (261 plus or minus 31 mM) and the difference in solium concentration between descending limb fluid and vasa recta was small and statistically insignificant. PMID:1127107

  20. Erythrocyte flow and dynamic hematocrit in the renal papilla of the rat.

    PubMed

    Zimmerhackl, B; Dussel, R; Steinhausen, M

    1985-12-01

    The microcirculation of the renal papilla was investigated in 32 vasa recta of Wistar rats. Using fluorescence microscopy in combination with a high-sensitivity television system we measured the velocity and flux of fluorescent-tagged erythrocytes in descending (DVR) and ascending vasa recta (AVR). After staining the plasma with fluorescent high molecular weight dextran we determined the diameters of DVR and AVR. Red cell flux (Qrbc) was determined from the ratio of the frequency of fluorescent-tagged red cells detected per unit time (fFITC) to the number of fluorescent-tagged red cells per nanoliter packed red cells (NFITC). From red cell velocity (Vrbc) and vessel diameter (D) we calculated the volume flow (Vapp). The dynamic hematocrit was directly derived as the ratio of Qrbc to Vapp. During antidiuresis Vrbc was 1.35 +/- 0.15 mm X s-1 (mean +/- SE) in DVR and 0.47 +/- 0.07 mm X s-1 in AVR. Qrbc in the same vessels averaged 3.26 +/- 0.9 and 1.72 +/- 0.35 nl X min-1, respectively. The diameter in DVR was 14.3 +/- 0.9 and in AVR 17.9 +/- 0.9 micron. From these values we calculated a dynamic hematocrit of 26 +/- 4 in DVR and 25 +/- 4% in AVR. The systemic hematocrit was 44 +/- 1%. The dynamic hematocrit in vasa recta represented 59 +/- 9 and 57 +/- 8% of the value in the systemic circulation, respectively.

  1. Hidden in the crowd: primordial germ cells and somatic stem cells in the mesodermal posterior growth zone of the polychaete Platynereis dumerillii are two distinct cell populations

    PubMed Central

    2012-01-01

    Background In the polychaete Platynereis, the primordial germ cells (PGCs) emerge from the vasa, piwi, and PL10 expressing mesodermal posterior growth zone (MPGZ) at the end of larval development, suggesting a post-embryonic formation from stem cells. Methods In order to verify this hypothesis, embryos and larvae were pulse labeled with the proliferation marker 5-ethynyl-2'-deoxyuridine (EdU) at different stages of development. Subsequently, the PGCs were visualized in 7-day-old young worms using antibodies against the Vasa protein. Results Surprisingly, the primordial germ cells of Platynereis incorporate EdU only shortly before gastrulation (6-8 hours post fertilization (hpf)), which coincides with the emergence of four small blastomeres from the mesoblast lineage. We conclude that these so-called 'secondary mesoblast cells' constitute the definitive PGCs in Platynereis. In contrast, the cells of the MPGZ incorporate EdU only from the pre-trochophore stage onward (14 hpf). Conclusion While PGCs and the cells of the MPGZ in Platynereis are indistinguishable in morphology and both express the germline markers vasa, nanos, and piwi, a distinct cluster of PGCs is detectable anterior of the MPGZ following EdU pulse-labeling. Indeed the PGCs form independently from the stem cells of the MPGZ prior to gastrulation. Our data suggest an early PGC formation in the polychaete by preformation rather than by epigenesis. PMID:22512981

  2. Regulation of hemopoiesis: inhibitors and stimulators produced by a murine bone marrow stromal cell line (H-1)

    SciTech Connect

    Cronkite, E.P.; Miller, M.E.; Garnett, H.; Harigaya, K.

    1982-01-01

    A murine cell line (H-1) probably derived from the adventitial reticular cell has been isolated and preserved. This line produces: (1) CSF; (2) a labile inhibitor of CFU-c proliferation with preferential action on granulopoiesis; (3) PGE; (4) proliferation inhibitors of BFU-E and GEMM; and (5) suppression of entry of CFU-S into DNA synthesis in vitro. It is postulated that the adventitial reticular cell (ARC) may play a major regulatory role in hemopoiesis through intramedullary production of the factors described. The nature of the signals that activate the genes in the ARC which are coded for the factors described is not known.

  3. Maize root characteristis that enhance flooding tolerance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant root systems have several cellular and molecular adaptations that are important in reducing stress caused by flooding. Of these, two physical properties of root systems provide an initial barrier toward the avoidance of stress. These are the presence of aerenchyma cells and rapid adventitious ...

  4. Imaging popliteal artery disease in young adults with claudication: self-assessment module.

    PubMed

    Chew, Felix S; Bui-Mansfield, Liem T

    2007-09-01

    The educational objectives of this self-assessment module on imaging popliteal artery disease in young adults with intermittent claudication are for the participant to exercise, self-assess, and improve his or her knowledge of the imaging and clinical features of popliteal artery entrapment syndrome, cystic adventitial disease,and masses associated with popliteal artery obstruction.

  5. Generating Leading Practices through Professional Learning

    ERIC Educational Resources Information Center

    Edwards Groves, Christine; Ronnerman, Karin

    2013-01-01

    In this paper we show how practices of professional learning and practices of leading can be understood as related in ecologies of practices. We will present findings from an international empirical research project that directs us to the connectivity between professional learning and leading practices that emerged as "adventitious",…

  6. Effect of polyvinyl alcohol on in vitro rooting capacity of shoots in pear clones (Pyrus communis L.) of different ploidy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poor adventitious root formation is a major obstacle in micropropagation. In this study, intense efforts have been made for improvement of rooting procedures for triploid, tetraploid, and mixploid clones of the pear cultivar, 'Fertility', obtained by in vitro colchicine treatment. An efficient roo...

  7. [Induction and in vitro culture of hairy roots of Dianthus caryophyllus and its plant regeneration].

    PubMed

    Shi, Heping; Zhu, Yuanfeng; Wang, Bei; Sun, Jiangbing; Huang, Shengqin

    2014-11-01

    To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.

  8. Personality Factors Associated with the Decision to Accept or Reject Mobility Training.

    ERIC Educational Resources Information Center

    White, K.; And Others

    1990-01-01

    Adventitiously blind adults (n=79) who had accepted mobility training were compared to 60 subjects who had rejected training. Personality profiles varied significantly between groups on seven scales of the Minnesota Multiphasic Personality Inventory: psychasthenia; schizophrenia; psychopathic deviate; depression; hypomania; paranoia; and…

  9. [Experiment on polyploid induction of Angelica dahurica var. formosana].

    PubMed

    Peng, F; Zhou, R; Liu, J

    1999-12-01

    Colchicine solution was applied to the primary adventitious buds of Angelica dahurica var. formosana in vitro to induce the polyploid. Compared with non-treated plantlet, the morphology, microhisology, and chromosome number of treated plantlets are varied. It proved that the polyploid induction was effective. PMID:12571900

  10. An efficient, widely applicable cryopreservation of Lilium shoot tips by droplet vitrification

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report a straightforward and widely applicable cryopreservation method for Lilium shoot tips. This method uses adventitious shoots that were induced from leaf segments cultured for 4 weeks on a shoot regeneration medium containing 1 mg L-1 a-naphthaleneacetic acid (NAA) and 0.5 mg L-1 thidiazuron...

  11. Developmental anatomy of blueberry (Vaccinium corymbosum L. ‘Aurora’) shoot regeneration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The culture of Vaccinium corymbosum L. ’Aurora’ leaves on regeneration medium results in the regeneration of adventitious shoots. We present anatomical evidence that these new shoot apices are directly regenerated from the cultured blades. Mounds of densely staining cells, which formed from epidermi...

  12. 7 CFR 201.56-11 - Knotweed family, Polygonaceae.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots developing within... tissue. (ii) Malformed, such as markedly shortened, curled, or thickened. (iii) Watery. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots....

  13. 7 CFR 201.56-11 - Knotweed family, Polygonaceae.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots developing within... tissue. (ii) Malformed, such as markedly shortened, curled, or thickened. (iii) Watery. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots....

  14. 7 CFR 201.56-11 - Knotweed family, Polygonaceae.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots developing within... tissue. (ii) Malformed, such as markedly shortened, curled, or thickened. (iii) Watery. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots....

  15. 7 CFR 201.56-8 - Flax family, Linaceae.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots usually developing... markedly shortened, curled, or thickened. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots. (5) Seedling: (i) One or more essential structures...

  16. 7 CFR 201.56-11 - Knotweed family, Polygonaceae.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots developing within... tissue. (ii) Malformed, such as markedly shortened, curled, or thickened. (iii) Watery. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots....

  17. 7 CFR 201.56-8 - Flax family, Linaceae.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots usually developing... markedly shortened, curled, or thickened. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots. (5) Seedling: (i) One or more essential structures...

  18. 7 CFR 201.56-8 - Flax family, Linaceae.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots usually developing... markedly shortened, curled, or thickened. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots. (5) Seedling: (i) One or more essential structures...

  19. 7 CFR 201.56-8 - Flax family, Linaceae.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots usually developing... markedly shortened, curled, or thickened. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots. (5) Seedling: (i) One or more essential structures...

  20. 7 CFR 201.56-11 - Knotweed family, Polygonaceae.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots developing within... tissue. (ii) Malformed, such as markedly shortened, curled, or thickened. (iii) Watery. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots....

  1. 7 CFR 201.56-8 - Flax family, Linaceae.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... development within the test period. (4) Root system: A primary root, with secondary roots usually developing... markedly shortened, curled, or thickened. (4) Root: (i) None. (ii) Weak, stubby, or missing primary root with weak secondary or adventitious roots. (5) Seedling: (i) One or more essential structures...

  2. Origin of Matrix-Producing Cells That Contribute to Aortic Fibrosis in Hypertension.

    PubMed

    Wu, Jing; Montaniel, Kim Ramil C; Saleh, Mohamed A; Xiao, Liang; Chen, Wei; Owens, Gary K; Humphrey, Jay D; Majesky, Mark W; Paik, David T; Hatzopoulos, Antonis K; Madhur, Meena S; Harrison, David G

    2016-02-01

    Various hypertensive stimuli lead to exuberant adventitial collagen deposition in large arteries, exacerbating blood pressure elevation and end-organ damage. Collagen production is generally attributed to resident fibroblasts; however, other cells, including resident and bone marrow-derived stem cell antigen positive (Sca-1(+)) cells and endothelial and vascular smooth muscle cells, can produce collagen and contribute to vascular stiffening. Using flow cytometry and immunofluorescence, we found that adventitial Sca-1(+) progenitor cells begin to produce collagen and acquire a fibroblast-like phenotype in hypertension. We also found that bone marrow-derived cells represent more than half of the matrix-producing cells in hypertension, and that one-third of these are Sca-1(+). Cell sorting and lineage-tracing studies showed that cells of endothelial origin contribute to no more than one fourth of adventitial collagen I(+) cells, whereas those of vascular smooth muscle lineage do not contribute. Our findings indicate that Sca-1(+) progenitor cells and bone marrow-derived infiltrating fibrocytes are major sources of arterial fibrosis in hypertension. Endothelial to mesenchymal transition likely also contributes, albeit to a lesser extent and pre-existing resident fibroblasts represent a minority of aortic collagen-producing cells in hypertension. This study shows that vascular stiffening represents a complex process involving recruitment and transformation of multiple cells types that ultimately elaborate adventitial extracellular matrix.

  3. Phytohormone balance and stress-related cellular responses are involved in the transition from bud to shoot growth in leafy spurge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Leafy spurge (Euphorbia esula L.) is an herbaceous weed that maintains a perennial growth habit through seasonal production of abundant underground adventitious buds (UABs) on the crown and lateral roots. During the normal growing season, differentiation of bud to shoot growth is inhibit...

  4. Glyphosate’s impact on vegetative growth in leafy spurge identifies molecular processes and hormone cross-talk associated with increased branching

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge (Euphorbia esula) is a perennial weed that is considered glyphosate tolerant. Tolerance is partly attributed to escape through establishment of new vegetative shoots from an abundance of underground adventitious buds. Sub-lethal concentrations of foliar applied glyphosate resulted in ne...

  5. Meta-analysis identifies potential molecular markers for endodormancy in crown buds of leafy spurge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vegetative shoot growth originating from underground adventitious buds (UABs) of herbaceous perennials such as leafy spurge (Euphorbia esula L.) is critical for survival after episodes of severe abiotic stress. Although leafy spurge is considered an invasive weed in North American ecosystems, it ha...

  6. Field application of glyphosate induces molecular changes affecting vegetative growth processes in leafy spurge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recommended rates of glyphosate for non-cultivated areas destroy the aboveground shoots of the perennial plant leafy spurge. However, such applications cause little or no damage to underground adventitious buds (UABs), and thus the plant readily regenerates vegetatively. High concentrations of glyph...

  7. Induction of endodormancy in crown buds of leafy spurge (Euphorbia esula L.) implicates a role for ethylene and cross-talk between photoperiod and temperature

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge is a model for studying well-defined phases of dormancy in underground adventitious buds (UABs) of herbaceous perennial weeds, which is a primary factor facilitating their escape from conventional control measures. A 12-week ramp down in both temperature (27°C ' 10°C) and photoperiod (1...

  8. Dehydration-induced endodormancy in crown buds of leafy spurge highlights involvement of MAF3- and RVE1-like homologs, and hormone signaling cross-talk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vegetative shoot growth from underground adventitious buds of leafy spurge is critical for survival of this invasive perennial weed after episodes of severe abiotic stress. To determine the impact that dehydration-stress has on molecular mechanisms associated with vegetative reproduction of leafy sp...

  9. Dehydration and vernalization treatments identify overlapping molecular networks impacting endodormancy maintenance in leafy spurge crown buds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge (Euphorbia esula L.) is an herbaceous perennial weed that reproduces vegetatively from an abundance of underground adventitious buds (UABs), which undergo well-defined phases of seasonal dormancy (para-, endo- and eco-dormancy). In this study, the effects of dehydration-stress on vegeta...

  10. Arterial mechanics considering the structural and mechanical contributions of ECM constituents.

    PubMed

    Wang, Yunjie; Zeinali-Davarani, Shahrokh; Zhang, Yanhang

    2016-08-16

    Considering the organization and engagement behavior of different extracellular matrix (ECM) constituents in the medial and adventitial layer of the arterial wall, in this study, we proposed a new constitutive model of ECM mechanics that considers the distinct structural and mechanical contributions of medial elastin, medial collagen, and adventitial collagen, to incorporate the constituent-specific fiber orientation and the sequential fiber engagement in arterial mechanics. Planar biaxial tensile testing method was used to characterize the orthotropic and hyperelastic behavior of porcine thoracic aorta. Fiber distribution functions of medial elastin, medial collagen, and adventitial collagen were incorporated into the constitutive model. Considering the sequential engagement of ECM constituents in arterial mechanics, a recruitment density function was incorporated into the model to capture the delayed engagement of adventitial collagen. A freely jointed chain model was used to capture the mechanical behavior of elastin and collagen at the fiber level. The tissue-level ECM mechanics was obtained by incorporating fiber distribution, engagement, and elastin and collagen content. The multi-scale constitutive model considering the structural and mechanical contributions of the three major ECM constituents allows us to directly incorporate information obtained from quantitative multi-photon imaging and analysis, and biochemical assay for the prediction of tissue-level mechanical response. Moreover, the model shows promises in fitting and predicting with a small set of material parameters, which has physical meanings and can be related to the structure of the ECM.

  11. Potential Bud Bank Responses to Apical Meristem Damage and Environmental Variables: Matching or Complementing Axillary Meristems?

    PubMed Central

    Klimešová, Jitka; Malíková, Lenka; Rosenthal, Jonathan; Šmilauer, Petr

    2014-01-01

    Soil nutrients, dormant axillary meristem availability, and competition can influence plant tolerance to damage. However, the role of potential bud banks (adventitious meristems initiated only after injury) is not known. Examining Central European field populations of 22 species of short-lived monocarpic herbs exposed to various sources of damage, we hypothesized that: (1) with increasing injury severity, the number of axillary branches would decrease, due to axillary meristem limitation, whereas the number of adventitious shoots (typically induced by severe injury) would increase; (2) favorable environmental conditions would allow intact plants to branch more, resulting in stronger axillary meristem limitation than in unfavorable conditions; and (3) consequently, adventitious sprouting would be better enabled in favorable than unfavorable conditions. We found strong support for the first hypothesis, only limited support for the second, and none for the third. Our results imply that whereas soil nutrients and competition marginally influence plant tolerance to damage, potential bud banks enable plants to overcome meristem limitation from severe damage, and therefore better tolerate it. All the significant effects were found in intraspecific comparisons, whereas interspecific differences were not found. Monocarpic plants with potential bud banks therefore represent a distinct strategy occupying a narrow environmental niche. The disturbance regime typical for this niche remains to be examined, as do the costs associated with the banks of adventitious and axillary reserve meristems. PMID:24516587

  12. Potential bud bank responses to apical meristem damage and environmental variables: matching or complementing axillary meristems?

    PubMed

    Klimešová, Jitka; Malíková, Lenka; Rosenthal, Jonathan; Šmilauer, Petr

    2014-01-01

    Soil nutrients, dormant axillary meristem availability, and competition can influence plant tolerance to damage. However, the role of potential bud banks (adventitious meristems initiated only after injury) is not known. Examining Central European field populations of 22 species of short-lived monocarpic herbs exposed to various sources of damage, we hypothesized that: (1) with increasing injury severity, the number of axillary branches would decrease, due to axillary meristem limitation, whereas the number of adventitious shoots (typically induced by severe injury) would increase; (2) favorable environmental conditions would allow intact plants to branch more, resulting in stronger axillary meristem limitation than in unfavorable conditions; and (3) consequently, adventitious sprouting would be better enabled in favorable than unfavorable conditions. We found strong support for the first hypothesis, only limited support for the second, and none for the third. Our results imply that whereas soil nutrients and competition marginally influence plant tolerance to damage, potential bud banks enable plants to overcome meristem limitation from severe damage, and therefore better tolerate it. All the significant effects were found in intraspecific comparisons, whereas interspecific differences were not found. Monocarpic plants with potential bud banks therefore represent a distinct strategy occupying a narrow environmental niche. The disturbance regime typical for this niche remains to be examined, as do the costs associated with the banks of adventitious and axillary reserve meristems. PMID:24516587

  13. Using Constant Time Delay to Teach Braille and the Nemeth Code for Mathematics and Science Notation to Students Making the Transition from Print to Braille

    ERIC Educational Resources Information Center

    Ivy, Sarah E.; Hooper, Jonathan D.

    2015-01-01

    Introduction: Many students with adventitious vision loss or progressive vision loss need to transition from print to braille as a primary literacy medium. It is important that this transition is handled efficiently so that the student can have continued access to a literacy medium and make progress in the core curriculum. For this study, we used…

  14. Bud Dormancy and Growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nearly all land plants produce ancillary meristems in the form of axillary or adventitious buds in addition to the shoot apical meristem. Outgrowth of these buds has a significant impact on plant architecture and the ability of plants to compete with neighboring plants, as well as to respond to and ...

  15. A simple method suitable to study de novo root organogenesis.

    PubMed

    Chen, Xiaodong; Qu, Yuliang; Sheng, Lihong; Liu, Jingchun; Huang, Hai; Xu, Lin

    2014-01-01

    De novo root organogenesis is the process in which adventitious roots regenerate from detached or wounded plant tissues or organs. In tissue culture, appropriate types and concentrations of plant hormones in the medium are critical for inducing adventitious roots. However, in natural conditions, regeneration from detached organs is likely to rely on endogenous hormones. To investigate the actions of endogenous hormones and the molecular mechanisms guiding de novo root organogenesis, we developed a simple method to imitate natural conditions for adventitious root formation by culturing Arabidopsis thaliana leaf explants on B5 medium without additive hormones. Here we show that the ability of the leaf explants to regenerate roots depends on the age of the leaf and on certain nutrients in the medium. Based on these observations, we provide examples of how this method can be used in different situations, and how it can be optimized. This simple method could be used to investigate the effects of various physiological and molecular changes on the regeneration of adventitious roots. It is also useful for tracing cell lineage during the regeneration process by differential interference contrast observation of β-glucuronidase staining, and by live imaging of proteins labeled with fluorescent tags. PMID:24860589

  16. Update on RR gene flow research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The United States is a major exporter of alfalfa seed and hay and the organic dairy industry is one of the fastest growing agricultural sectors. Because of this, many alfalfa producers are impacted by market sensitivity to adventitious presence (AP) of GM traits. Our objective was to quantify the ex...

  17. Autophosphorylation affects protein complex formation and activity of CDK-activating kinase (Ee;CDKF;1) in leafy spurge (Euphorbia esula)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge is a deep rooted perennial weed that propagates both by seeds and underground adventitious buds located on the crown and roots (crown and root buds). As buds develop during the normal growing season, they are maintained in a quiescent state through correlative inhibition. To enhance our...

  18. CDK-activating kinase (Ee;CDKF;1) of leafy spurge (Euphorbia esula) forms both homo-dimers and homo-trimers in its native state

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge is a deep rooted perennial weed that propagates both by seeds and underground adventitious buds located on the crown and roots (crown and root buds). As buds develop during the normal growing season, they are maintained in a quiescent state through correlative inhibition. To enhance our...

  19. Early Experience and Spatial Functioning in the Blind.

    ERIC Educational Resources Information Center

    Schmitt, Terry Lyndell; Warren, David H.

    1982-01-01

    Superior performance by adventitiously blind Ss relative to congenitally blind Ss appeared on tasks where the stimuli were complex, familiar and/or apprehenced with arm's length of the body, but not on tasks where the stimuli were relatively simple, novel, and/or apprehenced only through locomotion. (Author/MC)

  20. Early Experience and Spatial Functioning in the Blind.

    ERIC Educational Resources Information Center

    Schmitt, Terry Lyndell

    A review of the literature on the spatial functioning of the blind, the results of a questionnaire of 75 sighted college students and their parents, and results of a study of 87 blind and seeing adults' performance on three spatial tasks are presented. The review suggested that adventitiously blind individuals perform better than congenitally…

  1. Aerenchyma and an inducible barrier to radial oxygen loss facilitate root aeration in upland, paddy and deep-water rice (Oryza sativa L.).

    PubMed

    Colmer, T D

    2003-01-01

    The present study evaluated waterlogging tolerance, root porosity and radial O(2) loss (ROL) from the adventitious roots, of seven upland, three paddy, and two deep-water genotypes of rice (Oryza sativa L.). Upland types, with the exception of one genotype, were as tolerant of 30 d soil waterlogging as the paddy and deep-water types. In all but one of the 12 genotypes, the number of adventitious roots per stem increased for plants grown in waterlogged, compared with drained, soil. When grown in stagnant deoxygenated nutrient solution, genotypic variation was evident for root porosity and rates of ROL, but there was no overall difference between plants from the three cultural types. Adventitious root porosity increased from 20-26 % for plants grown in aerated solution to 29-41 % for plants grown in stagnant solution. Growth in stagnant solution also induced a 'tight' barrier to ROL in the basal regions of adventitious roots of five of the seven upland types, all three paddy types, and the two deep-water types. The enhanced porosity provided a low resistance pathway for O(2) movement to the root tip, and the barrier to ROL in basal zones would have further enhanced longitudinal O(2) diffusion towards the apex, by diminishing losses to the rhizosphere. The plasticity in root physiology, as described above, presumably contributes to the ability of rice to grow in diverse environments that differ markedly in soil waterlogging, such as drained upland soils as well as waterlogged paddy fields. PMID:12509350

  2. Causes of Deafness: Retrospection and Omens.

    ERIC Educational Resources Information Center

    Champie, Joan

    1996-01-01

    This study reviewed records of several American schools for the deaf in the 19th century concerning the stated causes of deafness given by parents. The high rate of adventitious deafness is noted. Stated causes are categorized into: fevers, inflammations, medicines/poisons, trauma, heat/cold, ear problems, nervous system problems, head/neck…

  3. Summary of breakout session f: facility and equipment decontamination strategies.

    PubMed

    Norwood, Laurie P; Suvarna, Kalavati

    2011-01-01

    CONFERENCE PROCEEDING Proceedings of the PDA/FDA Adventitious Viruses in Biologics: Detection and Mitigation Strategies Workshop in Bethesda, MD, USA; December 1-3, 2010 Guest Editors: Arifa Khan (Bethesda, MD), Patricia Hughes (Bethesda, MD) and Michael Wiebe (San Francisco, CA).

  4. Design of a novel molecular beacon: modification of the stem with artificially genetic alphabet.

    PubMed

    Sheng, Pinpin; Yang, Zunyi; Kim, Youngmi; Wu, Yanrong; Tan, Weihong; Benner, Steven A

    2008-11-01

    A molecular beacon that incorporates components of an artificially expanded genetic information system (Aegis) in its stem is shown not to be opened by unwanted stem invasion by adventitious standard DNA; this should improve the "darkness" of the beacon in real-world applications.

  5. Extended Low Temperature Impacts Dormancy Status, Flowering Competence, and Transcript Profiles in Crown Buds of Leafy Spurge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leafy spurge (Euphorbia esula) is an herbaceous perennial weed that reproduces vegetatively from an abundance of underground adventitious buds. In this study we report the effects of different growth conditions on vegetative reproduction and flowering competence, and determine molecular mechanisms a...

  6. PDA Cell Substrate Task Force: Emerging Technologies for Virus Detection Technical Document--A Progress Report.

    PubMed

    King, Kathryn; Wiebe, Michael

    2011-01-01

    CONFERENCE PROCEEDING Proceedings of the PDA/FDA Adventitious Viruses in Biologics: Detection and Mitigation Strategies Workshop in Bethesda, MD, USA; December 1-3, 2010 Guest Editors: Arifa Khan (Bethesda, MD), Patricia Hughes (Bethesda, MD) and Michael Wiebe (San Francisco, CA). PMID:22294602

  7. Demonstrations at School Level of the Effects of IAA

    ERIC Educational Resources Information Center

    Falk, Peter

    1973-01-01

    Describes demonstrations suitable for secondary school biology classes relating to the effects of the hormone IAA on plant growth. Demonstrations illustrate how hormone treatments affect stem elongation, callus formation, inhibition of axillary buds, stimulation of secondary growth and initiation of adventitious root development, root elongation,…

  8. Benign Visual Hallucinations, or "Phantom Vision" in Visually Impaired and Blind Persons.

    ERIC Educational Resources Information Center

    Needham, W. E.; Taylor, R. E.

    1992-01-01

    Benign visual hallucinations ("phantom vision") were examined in 2 studies, involving a total of 443 adventitiously blinded and sight-impaired veterans. In one study, unusual visual events were reported by 38.7 percent, with 30.6 percent reporting complex hallucinations. Causes of the hallucinations and their treatment are discussed. (Author/JDD)

  9. Characterization and expression of a Pinus radiata putative ortholog to the Arabidopsis SHORT-ROOT gene.

    PubMed

    Solé, Alicia; Sánchez, Conchi; Vielba, Jesús M; Valladares, Silvia; Abarca, Dolores; Díaz-Sala, Carmen

    2008-11-01

    We characterized a Pinus radiata D. Don putative ortholog to the Arabidopsis thaliana (L.) Heynh. SHORT--ROOT gene (AtSHR) and analyzed its expression in different organs during vegetative development and in response to exogenous auxin during adventitious rooting. The predicted protein sequence contained domains characteristic of the GRAS protein family and showed a strong similarity to the SHORT--ROOT (SHR) proteins. Quantitative reverse transcriptase--polymerase chain reaction (qRT-PCR) and in situ hybridization showed that the gene is predominantly expressed in roots, root primordia and in the cambial region of hypocotyl cuttings. Increased mRNA levels were observed, independently of the presence or absence of exogenous auxin, in the cambial region and rooting competent cells of hypocotyl cuttings within the first 24 h of adventitious rooting, before the activation of cell divisions and the organization of the adventitious root meristem. The expression pattern in organs and during adventitious rooting was similar to that of a Pinus radiata SCARECROW-LIKE (PrSCL1) gene, except that PrSCL1 is induced in response to exogenous auxin. Results suggest that the Pinus radiata SHORT-ROOT (PrSHR) gene has a role in root meristem formation and maintenance and in the cambial region of hypocotyl cuttings.

  10. [Establishment of high frequency regeneration via leaf explants of 'Red Sun' kiwifruit (Actinidia chinensis)].

    PubMed

    Zhao, Xupeng; Luo, Keming; Zhou, Yue; Wu, Xiuhua; Yang, Li; Tang, Shaohu

    2013-11-01

    A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit.

  11. Plant regeneration and genetic transformation of C. canadensis: a non-model plant appropriate for investigation of flower development in Cornus (Cornaceae).

    PubMed

    Liu, Xiang; Feng, Chun-Miao; Franks, Robert; Qu, Rongda; Xie, De-Yu; Xiang, Qiu-Yun Jenny

    2013-01-01

    KEY MESSAGE : Efficient Agrobacterium -mediated genetic transformation for investigation of genetic and molecular mechanisms involved in inflorescence architectures in Cornus species. Cornus canadensis is a subshrub species in Cornus, Cornaceae. It has recently become a favored non-model plant species to study genes involved in development and evolution of inflorescence architectures in Cornaceae. Here, we report an effective protocol of plant regeneration and genetic transformation of C. canadensis. We use young inflorescence buds as explants to efficiently induce calli and multiple adventitious shoots on an optimized induction medium consisting of basal MS medium supplemented with 1 mg/l of 6-benzylaminopurine and 0.1 mg/l of 1-naphthaleneacetic acid. On the same medium, primary adventitious shoots can produce a large number of secondary adventitious shoots. Using leaves of 8-week-old secondary shoots as explants, GFP as a reporter gene controlled by 35S promoter and hygromycin B as the selection antibiotic, a standard procedure including pre-culture of explants, infection, co-cultivation, resting and selection has been developed to transform C. canadensis via Agrobacterium strain EHA105-mediated transformation. Under a strict selection condition using 14 mg/l hygromycin B, approximately 5 % explants infected by Agrobacterium produce resistant calli, from which clusters of adventitious shoots are induced. On an optimized rooting medium consisting of basal MS medium supplemented with 0.1 mg/l of indole-3-butyric acid and 7 mg/l hygromycin B, most of the resistant shoots develop adventitious roots to form complete transgenic plantlets, which can grow normally in soil. RT-PCR analysis demonstrates the expression of GFP transgene. Green fluorescence emitted by GFP is observed in transgenic calli, roots and cells of transgenic leaves under both stereo fluorescence microscope and confocal microscope. The success of genetic transformation provides an appropriate

  12. Nanoparticle biofabrication using English ivy (Hedera helix)

    PubMed Central

    2012-01-01

    Background English ivy (Hedera helix) is well known for its adhesive properties and climbing ability. Essential to its ability to adhere to vertical surfaces is the secretion of a nanocomposite adhesive containing spherical nanoparticles, 60–85 nm in diameter, produced exclusively by root hairs present on adventitious roots. These organic nanoparticles have shown promise in biomedical and cosmetic applications, and represent a safer alternative to metal oxide nanoparticles currently available. Results It was discovered that the maximum adventitious root production was achieved by a 4 h application of 1 mg/ml indole-3 butyric acid (IBA) to juvenile English ivy shoot segments cultured in custom vessels. After incubation of the shoots under continuous light at 83 μmol/m2 s at 20°C for 2 weeks, the adventitious roots were harvested from the culture system and it was possible to isolate 90 mg of dry weight nanoparticles per 12 g of roots. The nanoparticle morphology was characterized by atomic force microscopy, and found to be similar to previous studies. Conclusions An enhanced system for the production of English ivy adventitious roots and their nanoparticles by modifying GA7 Magenta boxes and identifying the optimal concentration of IBA for adventitious root growth was developed. This system is the first such platform for growing and harvesting organic nanoparticles from plants, and represents an important step in the development of plant-based nanomanufacturing. It is a significant improvement on the exploitation of plant systems for the formation of metallic nanoparticles, and represents a pathway for the generation of bulk ivy nanoparticles for translation into biomedical applications. PMID:23095780

  13. Different Responses of an Invasive Clonal Plant Wedelia trilobata and its Native Congener to Gibberellin: Implications for Biological Invasion.

    PubMed

    Dai, Zhi-Cong; Fu, Wei; Qi, Shan-Shan; Zhai, De-Li; Chen, Si-Chong; Wan, Ling-Yun; Huang, Ping; Du, Dao-Lin

    2016-02-01

    The invasive clonal plant Wedelia trilobata contains higher levels of ent-kaurane diterpenes, which are precursors of gibberellins (GAs), and higher rates of clonal growth than its native congener W. chinensis in invaded habitats. We hypothesized that the higher levels of endogenous GAs facilitate greater ramet growth in W. trilobata compared with W. chinensis. We quantified endogenous levels of GA1+3 in the two species and compared their growth responses to the changes of endogenous and exogenous GA3 by using short-term and long-term hydroponics experiments. After a period of homogeneous cultivation, levels of endogenous GA1+3 were higher in W. trilobata than in W. chinensis. The reduction of endogenous GAs repressed the emergence of adventitious roots and the growth of W. trilobata in the initial cultivation stage, and inhibited its shoot elongation and biomass. Levels of endogenous GA1+3 were positively correlated with the length of shoots and adventitious roots of W. trilobata. Adventitious roots of W. trilobata also emerged earlier and grew faster when treated with exogenous GA3. In contrast, exogenous GA3 treatment inhibited the length of adventitious roots in W. chinensis, and levels of endogenous GA1+3 did not correlate with shoot or adventitious root length. Our study suggests that GAs accelerate the rapid clonal growth of W. trilobata, more than that of its native congener W. chinensis, illustrating the relationship between plant hormones and the clonal growth of invasive plants. These findings are important for understanding the mechanisms associated with the invasiveness of clonal plants and their potential management. PMID:26879680

  14. Perivascular cells for regenerative medicine

    PubMed Central

    Crisan, Mihaela; Corselli, Mirko; Chen, William CW; Péault, Bruno; Moldovan, NI

    2012-01-01

    Mesenchymal stem/stromal cells (MSC) are currently the best candidate therapeutic cells for regenerative medicine related to osteoarticular, muscular, vascular and inflammatory diseases, although these cells remain heterogeneous and necessitate a better biological characterization. We and others recently described that MSC originate from two types of perivascular cells, namely pericytes and adventitial cells and contain the in situ counterpart of MSC in developing and adult human organs, which can be prospectively purified using well defined cell surface markers. Pericytes encircle endothelial cells of capillaries and microvessels and express the adhesion molecule CD146 and the PDGFRβ, but lack endothelial and haematopoietic markers such as CD34, CD31, vWF (von Willebrand factor), the ligand for Ulex europaeus 1 (UEA1) and CD45 respectively. The proteoglycan NG2 is a pericyte marker exclusively associated with the arterial system. Besides its expression in smooth muscle cells, smooth muscle actin (αSMA) is also detected in subsets of pericytes. Adventitial cells surround the largest vessels and, opposite to pericytes, are not closely associated to endothelial cells. Adventitial cells express CD34 and lack αSMA and all endothelial and haematopoietic cell markers, as for pericytes. Altogether, pericytes and adventitial perivascular cells express in situ and in culture markers of MSC and display capacities to differentiate towards osteogenic, adipogenic and chondrogenic cell lineages. Importantly, adventitial cells can differentiate into pericyte-like cells under inductive conditions in vitro. Altogether, using purified perivascular cells instead of MSC may bring higher benefits to regenerative medicine, including the possibility, for the first time, to use these cells uncultured. PMID:22882758

  15. Different Responses of an Invasive Clonal Plant Wedelia trilobata and its Native Congener to Gibberellin: Implications for Biological Invasion.

    PubMed

    Dai, Zhi-Cong; Fu, Wei; Qi, Shan-Shan; Zhai, De-Li; Chen, Si-Chong; Wan, Ling-Yun; Huang, Ping; Du, Dao-Lin

    2016-02-01

    The invasive clonal plant Wedelia trilobata contains higher levels of ent-kaurane diterpenes, which are precursors of gibberellins (GAs), and higher rates of clonal growth than its native congener W. chinensis in invaded habitats. We hypothesized that the higher levels of endogenous GAs facilitate greater ramet growth in W. trilobata compared with W. chinensis. We quantified endogenous levels of GA1+3 in the two species and compared their growth responses to the changes of endogenous and exogenous GA3 by using short-term and long-term hydroponics experiments. After a period of homogeneous cultivation, levels of endogenous GA1+3 were higher in W. trilobata than in W. chinensis. The reduction of endogenous GAs repressed the emergence of adventitious roots and the growth of W. trilobata in the initial cultivation stage, and inhibited its shoot elongation and biomass. Levels of endogenous GA1+3 were positively correlated with the length of shoots and adventitious roots of W. trilobata. Adventitious roots of W. trilobata also emerged earlier and grew faster when treated with exogenous GA3. In contrast, exogenous GA3 treatment inhibited the length of adventitious roots in W. chinensis, and levels of endogenous GA1+3 did not correlate with shoot or adventitious root length. Our study suggests that GAs accelerate the rapid clonal growth of W. trilobata, more than that of its native congener W. chinensis, illustrating the relationship between plant hormones and the clonal growth of invasive plants. These findings are important for understanding the mechanisms associated with the invasiveness of clonal plants and their potential management.

  16. Fleas infesting pets in the era of emerging extra-intestinal nematodes

    PubMed Central

    2013-01-01

    Modifications in climatic conditions, movements of hosts and goods, changes in animal phenology and human behaviour and increase of wildlife, are presently concurring in the geographic spread of vectors and cardio-respiratory nematodes, e.g. Dirofilaria immitis, Angiostrongylus vasorum, Aelurostrongylus abstrusus and Capillaria aerophila. All these factors may also influence dispersion and clinical significance of fleas, thus posing relevant challenges in those regions where other parasites are emerging at the same time. Ctenocephalides felis, Ctenocephalides canis and Pulex irritans cause discomfort, nuisance, allergic reactions, anaemia, and may transmit several pathogens, some of them are of importance for public health. The present article reviews the importance of fleas in small animal practice and their sanitary relevance for dogs, cats and humans, and discusses current control methods in the present era of emerging extra-intestinal nematodes, towards a possible changing perspective for controlling key parasites affecting companion animals. PMID:23497511

  17. Canine and feline cardiopulmonary parasitic nematodes in Europe: emerging and underestimated

    PubMed Central

    2010-01-01

    Cardiopulmonary nematodes of dogs and cats cause parasitic diseases of central relevance in current veterinary practice. In the recent past the distribution of canine and feline heartworms and lungworms has increased in various geographical areas, including Europe. This is true especially for the metastrongyloids Aelurostrongylus abstrusus, Angiostrongylus vasorum and Crenosoma vulpis, the filarioid Dirofilaria immitis and the trichuroid Eucoleus aerophilus (syn. Capillaria aerophila). The reasons of this emergence are little known but many drivers such as global warming, changes in vector epidemiology and movements in animal populations, may be taken into account. The purpose of this article is to review the knowledge of the most important heartworm and lungworm infections of dogs and cats in Europe. In particular recent advances in epidemiology, clinical and control are described and discussed. PMID:20653938

  18. Bone morphogenetic protein 4 (BMP4) induces buffalo (Bubalus bubalis) embryonic stem cell differentiation into germ cells.

    PubMed

    Shah, Syed Mohmad; Saini, Neha; Ashraf, Syma; Singh, Manoj Kumar; Manik, Radhey Sham; Singla, Suresh Kumar; Palta, Prabhat; Chauhan, Manmohan Singh

    2015-12-01

    The aim of the present study was to investigate the effect of Bone morphogenetic protein 4 (BMP4) stimulation on differentiation of buffalo embryonic stem (ES) cells into germ lineage cells. ES cells were subjected to in vitro differentiation in floating and adherent cultures, under different BMP4 concentrations (20, 50 and 100 ngml(-1)) for different culture intervals (4, 8 and 14 days). qPCR analysis revealed that BMP4 at a concentration of 50-100 ngml(-1) for a culture period of 14 days led to maximum induction of germ lineage genes like DAZL, VASA, PLZF (PGC-specific); SYCP3, MLH1, TNP1/2 and PRM2 (Meiotic genes); BOULE and TEKT1 (Spermatocyte markers); GDF9, ZP2 and 3 (Oocyte markers). The expression levels of all the genes were significantly higher under BMP4 differentiation as compared to BMP4 + NOGGIN and spontaneously differentiated cultures. Immunocytochemical analysis of embryoid bodies (EBs) and monolayer adherent cultures revealed expression of PGC- (c-KIT, DAZL and VASA); Meiotic- (SYCP3, MLH1 and PROTAMINE1); Spermatocyte- (ACROSIN and HAPRIN); and Oocyte- markers (GDF9 and ZP4). Western blotting was positive for VASA, GDF9 and ZP4. Oocyte-like structures (OLS) obtained in monolayer differentiated cultures harbored a big nucleus and a ZP4 coat. They showed embryonic development and progressed through 2-cell, 4-cell, 8-cell and blastocyst-like structures. Global DNA methylation analysis showed significantly (p < 0.05) decreased levels of 5-methyl-2-deoxycytidine in EBs obtained in optimum differentiation medium. The expression of meiotic markers coupled with expression of spermatocyte and oocyte markers is an indication of post-meiotic progression into spermatogenesis and oogenesis, respectively.

  19. Tissue regeneration and biomineralization in sea urchins: role of Notch signaling and presence of stem cell markers.

    PubMed

    Reinardy, Helena C; Emerson, Chloe E; Manley, Jason M; Bodnar, Andrea G

    2015-01-01

    Echinoderms represent a phylum with exceptional regenerative capabilities that can reconstruct both external appendages and internal organs. Mechanistic understanding of the cellular pathways involved in regeneration in these animals has been hampered by the limited genomic tools and limited ability to manipulate regenerative processes. We present a functional assay to investigate mechanisms of tissue regeneration and biomineralization by measuring the regrowth of amputated tube feet (sensory and motor appendages) and spines in the sea urchin, Lytechinus variegatus. The ability to manipulate regeneration was demonstrated by concentration-dependent inhibition of regrowth of spines and tube feet by treatment with the mitotic inhibitor, vincristine. Treatment with the gamma-secretase inhibitor DAPT resulted in a concentration-dependent inhibition of regrowth, indicating that both tube feet and spine regeneration require functional Notch signaling. Stem cell markers (Piwi and Vasa) were expressed in tube feet and spine tissue, and Vasa-positive cells were localized throughout the epidermis of tube feet by immunohistochemistry, suggesting the existence of multipotent progenitor cells in these highly regenerative appendages. The presence of Vasa protein in other somatic tissues (e.g. esophagus, radial nerve, and a sub-population of coelomocytes) suggests that multipotent cells are present throughout adult sea urchins and may contribute to normal homeostasis in addition to regeneration. Mechanistic insight into the cellular pathways governing the tremendous regenerative capacity of echinoderms may reveal processes that can be modulated for regenerative therapies, shed light on the evolution of regeneration, and enable the ability to predict how these processes will respond to changing environmental conditions.

  20. Tissue Regeneration and Biomineralization in Sea Urchins: Role of Notch Signaling and Presence of Stem Cell Markers

    PubMed Central

    Reinardy, Helena C.; Emerson, Chloe E.; Manley, Jason M.; Bodnar, Andrea G.

    2015-01-01

    Echinoderms represent a phylum with exceptional regenerative capabilities that can reconstruct both external appendages and internal organs. Mechanistic understanding of the cellular pathways involved in regeneration in these animals has been hampered by the limited genomic tools and limited ability to manipulate regenerative processes. We present a functional assay to investigate mechanisms of tissue regeneration and biomineralization by measuring the regrowth of amputated tube feet (sensory and motor appendages) and spines in the sea urchin, Lytechinus variegatus. The ability to manipulate regeneration was demonstrated by concentration-dependent inhibition of regrowth of spines and tube feet by treatment with the mitotic inhibitor, vincristine. Treatment with the gamma-secretase inhibitor DAPT resulted in a concentration-dependent inhibition of regrowth, indicating that both tube feet and spine regeneration require functional Notch signaling. Stem cell markers (Piwi and Vasa) were expressed in tube feet and spine tissue, and Vasa-positive cells were localized throughout the epidermis of tube feet by immunohistochemistry, suggesting the existence of multipotent progenitor cells in these highly regenerative appendages. The presence of Vasa protein in other somatic tissues (e.g. esophagus, radial nerve, and a sub-population of coelomocytes) suggests that multipotent cells are present throughout adult sea urchins and may contribute to normal homeostasis in addition to regeneration. Mechanistic insight into the cellular pathways governing the tremendous regenerative capacity of echinoderms may reveal processes that can be modulated for regenerative therapies, shed light on the evolution of regeneration, and enable the ability to predict how these processes will respond to changing environmental conditions. PMID:26267358

  1. Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs.

    PubMed

    Nakamura, Yoriko; Iwasaki, Takehiro; Umei, Yosuke; Saotome, Kazuhiro; Nakajima, Yukiko; Kitahara, Shoichi; Uno, Yoshinobu; Matsuda, Yoichi; Oike, Akira; Kodama, Maho; Nakamura, Masahisa

    2015-10-01

    The Pat1 gene is expressed in the immature oocytes of Xenopus, and is reportedly involved in regulating the translation of maternal mRNAs required for oocyte-maturation. However, it is still unknown when Pat1a first appears in the differentiating ovary of amphibians. To address this issue, we isolated the full-length Pat1a cDNA from the frog Rana rugosa and examined its expression in the differentiating ovary of this frog. Among eight different tissues examined, the Pat1a mRNA was detectable in only the ovary. When frozen sections from the ovaries of tadpoles at various stages of development were immunostained for Vasa-a germ cell-specific protein-and Pat1a, Vasa-immunopositive signals were observed in all of the germ cells, whereas Pat1a signals were confined to the growing oocytes (50-200 μm in diameter), and absent from small germ cells (<50 μm in diameter). Forty days after testosterone injection into tadpoles to induce female-to-male sex-reversal, Pat1a-immunoreactive oocytes had disappeared completely from the sex-reversed gonad, but Vasa-positive small germ cells persisted. Thus, Pat1a would be a good marker for identifying the sexual status of the sex-reversing gonad in amphibians. In addition, fluorescence in situ hybridization analysis showed Pat1a to have an autosomal locus, suggesting that Pat1a transcription is probably regulated by a tissue-specific transcription factor in R. rugosa.

  2. Cytodifferentiation during the spermatogenesis of the hermaphrodite caridea Exhippolysmata oplophoroides.

    PubMed

    Nunes, Erika Takagi; Braga, Adriane Araújo; dos Santos, Daniela Carvalho; Camargo-Mathias, Maria Izabel

    2010-08-01

    Among the decapods, the caridean Exhippolysmata oplophoroides has been described as a simultaneous protandric hermaphrodite, seeing that it presents a male initial stage followed by a hermaphrodite one in which it can function as male and as female. This work had the aims of characterizing the microscopical morphology of the male portion of the ovotestes gonads from E. oplophoroides, at the different development stages, identifying each cell from the germ lines during spermatogenesis, as well as describing the ultramorphology of spermatozoans in the terminal region of the vasa deferentia. Shrimps were collected in Ubatuba, north coast of São Paulo, and their male gonads and the ampoule were removed, fixed and processed according to histological routine and for scanning electron microscopy. The testicular portion is divided in lobes, inside which cells at the same stage of the spermatogenic cycle are observed, with prevalence of spermatogonia and spermatocytes at stages I, II and V of gonad development, whereas spermatids and spermatozoans are found at stages III and IV, respectively. Ultramorphology of the terminal portion of the vasa deferentia exhibits mature aflagellated spike-shaped spermatozoans, encased in secretion and between membrane foldings that will constitute the spermatophores. Despite presenting reproductive characteristics common to other decapods, E. oplophoroides shows spermatozoans as well as spermatophore with typical morphology, which is important for its identification and taxonomy. Further, this species showed polysaccharide secretions where the spermatozoa are immerse as far as the testicular portion, which could have a role in their transport and nutrition as well as spermatophore constitution and/or fixation; differently, other caridean species begin spermatophore formation during the passage of the gametes through the vasa deferentia.

  3. Differential expression of T- and L-type voltage-dependent calcium channels in renal resistance vessels.

    PubMed

    Hansen, P B; Jensen, B L; Andreasen, D; Skøtt, O

    2001-09-28

    The distribution of voltage-dependent calcium channels in kidney pre- and postglomerular resistance vessels was determined at the molecular and functional levels. Reverse transcription-polymerase chain reaction analysis of microdissected rat preglomerular vessels and cultured smooth muscle cells showed coexpression of mRNAs for T-type subunits (Ca(V)3.1, Ca(V)3.2) and for an L-type subunit (Ca(V)1.2). The same expression pattern was observed in juxtamedullary efferent arterioles and outer medullary vasa recta. No calcium channel messages were detected in cortical efferent arterioles. Ca(V)1.2 protein was demonstrated by immunochemical labeling of rat preglomerular vasculature and juxtamedullary efferent arterioles and vasa recta. Cortical efferent arterioles were not immunopositive. Recordings of intracellular calcium concentration with digital fluorescence imaging microscopy showed a significant increase of calcium in response to K(+) (100 mmol/L) in isolated afferent arterioles (140+/-25%) and in juxtamedullary efferent arterioles (118+/-21%). These calcium responses were attenuated by the L-type antagonist calciseptine and by the T-type antagonist mibefradil. Intracellular calcium increased in response to K(+) in cortical efferent arterioles (21+/-9%). Mibefradil and nickel concentration dependently blocked K(+)-induced contraction of perfused rabbit afferent arterioles. Calciseptine blocked the contraction mediated by K(+) (EC(50) 8x10(-14)). S-(-)-Bay K 8644 had no effect on vascular diameter in the afferent arteriole. We conclude that voltage-dependent L- and T-type calcium channels are expressed and of functional significance in renal cortical preglomerular vessels, in juxtamedullary efferent arterioles, and in outer medullary vasa recta, but not in cortical efferent arterioles.

  4. Interaction of Xylamine with peripheral sympathetic neurons

    SciTech Connect

    Ransom, R.W.; Waggaman, L.A.; Cho, A.K.

    1985-09-30

    Xylamine (XYL) administered to intact rats caused a 70-80% reduction in norepinephrine (NE) uptake by the vas deferens but had little or no effect on NE content in that tissue. The vas deferens accumulates /sup 3/H-XYL in vitro by a desmethylimipramine (DMI)-sensitive mechanism. Vasa deferentia from 6-hydroxydopamine (60HDA) pretreated animals exhibited a 80% reduction in both NE content and XYL uptake activity. These results indicate that XYL is taken up by sympathetic nerve terminals and can reduce NE uptake activity without depleting terminals of neurotransmitter. 9 references, 4 tables.

  5. One Thousand Vasectomies

    PubMed Central

    Barnes, Marian N.; Blandy, J. P.; England, H. R.; Gunn, George; Howard, Geraldine; Law, Barbara; Mason, Bridgett; Medawar, Jean; Reynolds, C.; Shearer, R. J.; Singh, M.; Stanley-Roose, Dorothy G.

    1973-01-01

    In a prospective study of 1,000 people treated by vasectomy at the Margaret Pyke Centre under local anaesthesia as outpatients minor sepsis occurred in 12 cases, an abscess in one, and haematoma needing drainage in seven. Spontaneous recanalization of the vasa occurred in six patients and an unsuspected third vas was found in one. Except in patients with recanalization or a third vas the semen was always free from motile spermatozoa within three months, but non-motile spermatozoa have so far persisted for up to 17 months in a few cases. PMID:4758395

  6. Effects of denervation and reserpine on nexuses in the rat vas deferens.

    PubMed

    Westfall, D P; Millecchia, L L; Lee, T J; Corey, S P; Smith, D J; Fleming, W W

    1977-01-21

    Permanganate-fixed vasa deferentia from rats were examined for the presence of nexal-like contacts by electron microscopy. There was a significantly greater incidence of nexuses (2X) in chronically denervated tissues (5-7 days) but not in tissues from reserpine-pretreated animals (1.0 mg/kg/day for 5-7 days). The results suggest that an increase in nexal regions may not be a general feature of postjunctional supersensitivity but rather may contribute to other denervation-induced changes in contractile response. PMID:832678

  7. A conserved germline multipotency program

    PubMed Central

    Juliano, Celina E.; Swartz, S. Zachary; Wessel, Gary M.

    2010-01-01

    The germline of multicellular animals is segregated from somatic tissues, which is an essential developmental process for the next generation. Although certain ecdysozoans and chordates segregate their germline during embryogenesis, animals from other taxa segregate their germline after embryogenesis from multipotent progenitor cells. An overlapping set of genes, including vasa, nanos and piwi, operate in both multipotent precursors and in the germline. As we propose here, this conservation implies the existence of an underlying germline multipotency program in these cell types that has a previously underappreciated and conserved function in maintaining multipotency. PMID:21098563

  8. Hedgehog and Resident Vascular Stem Cell Fate

    PubMed Central

    Mooney, Ciaran J.; Hakimjavadi, Roya; Fitzpatrick, Emma; Kennedy, Eimear; Walls, Dermot; Morrow, David; Redmond, Eileen M.; Cahill, Paul A.

    2015-01-01

    The Hedgehog pathway is a pivotal morphogenic driver during embryonic development and a key regulator of adult stem cell self-renewal. The discovery of resident multipotent vascular stem cells and adventitial progenitors within the vessel wall has transformed our understanding of the origin of medial and neointimal vascular smooth muscle cells (SMCs) during vessel repair in response to injury, lesion formation, and overall disease progression. This review highlights the importance of components of the Hh and Notch signalling pathways within the medial and adventitial regions of adult vessels, their recapitulation following vascular injury and disease progression, and their putative role in the maintenance and differentiation of resident vascular stem cells to vascular lineages from discrete niches within the vessel wall. PMID:26064136

  9. Vegetative propagation of Cecropia obtusifolia (Cecropiaceae).

    PubMed

    LaPierre, L M

    2001-01-01

    Cecropia is a relatively well-known and well-studied genus in the Neotropics. Methods for the successful propagation of C. obtusifolia Bertoloni, 1840 from cuttings and air layering are described, and the results of an experiment to test the effect of two auxins, naphthalene acetic acid (NAA) and indole butyric acid (IBA), on adventitious root production in cuttings are presented. In general, C. obtusifolia cuttings respond well to adventitious root production (58.3% of cuttings survived to root), but air layering was the better method (93% of cuttings survived to root). The concentration of auxins used resulted in an overall significantly lower quality of roots produced compared with cuttings without auxin treatment. Future experiments using Cecropia could benefit from the use of isogenic plants produced by vegetative propagation. PMID:12189829

  10. An Efficient Protocol for Plantlet Regeneration via Direct Organogenesis by Using Nodal Segments from Embryo-Cultured Seedlings of Cinnamomum camphora L.

    PubMed Central

    Du, Li; Li, Yongpeng; Yao, Yao; Zhang, Liwei

    2015-01-01

    A simple and efficient plantlet regeneration protocol via direct organogenesis was established for camphor tree (Cinnamomum camphora L.). Stem segments with one node (SN explants) from embryo-cultured seedlings (EC seedlings) were used as explants. Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2, 4-dichlorophenoxyacetic acid and 2.0 mg/L 6-benzyladenine was used to induce cotyledonary embryo germination. This medium was also used for EC seedlings propagation and adventitious bud induction from SN explants. Regenerated plantlets were cultured on hormone-free MS medium for elongation and root induction. The regeneration capability of SN explants was compared by using EC seedling lines established in this research. EC seedling line EL6 exhibited the highest adventitious bud induction frequency (91.7%) and the highest number of buds per responding explant (5.2), which was considered as the most efficient EC seedling line for further gene transformation research. PMID:25962170

  11. Hallux valgus and hallux rigidus: MRI findings.

    PubMed

    Schweitzer, M E; Maheshwari, S; Shabshin, N

    1999-01-01

    The purpose of this article is to describe the MR findings of Hallux Valgus (HV) and Hallux Rigidus (HR). Twenty-four patients (11 with HV, 4 with HR, and 9 with both HV and HR) were studied at 1.5 Tesla MRI. Two separate observers evaluated the first ray blindly for the following signs: sesamoid position, sesamoid proliferation, hypertrophy of the median eminence, presence of a lateral facet, presence of an adventitial bursa, shape of the first metatarsal head, relative length of the first metatarsal, joint space loss, osteophytes (dorsalor lateral), marrow edema, geodes, subchondral sclerosis, intra-articular ossicle, and pes planus. The most common findings observed in HV were a hypertrophic medial eminence (95%), sesamoid proliferation (90%) and adventitial bursitis (70%). The most common findings observed in HR were osteophytes (77% and 69%), geodes, and marrow edema. We conclude that traditional routine radiograph signs of HV and HR may be applied to MR images.

  12. Regulating coexistence of GM and non-GM crops without jeopardizing economic incentives.

    PubMed

    Demont, Matty; Devos, Yann

    2008-07-01

    The ongoing debate about the coexistence of genetically modified (GM) and non-GM crops in the European Union (EU) mainly focuses on preventive measures needed to keep the adventitious presence of GM material in non-GM products below established tolerance thresholds, as well as on issues covering questions of liability and the duty to redress the incurred economic harm once adventitious mixing in non-GM products has occurred. By contrast, the interplay between the economic incentives and costs of coexistence has attracted little attention. The current overemphasis on the technical aspects and cost of coexistence over its economic incentives might lead EU policy-makers to adopt too stringent and rigid regulations on coexistence. Therefore, we argue for flexible coexistence regulations that explicitly take into account the economic incentives for coexistence. Our arguments provide a timely and important framework for EU policy-makers, who are currently struggling to implement coherent coexistence regulations in all member states.

  13. Influence of aluminium on the restoration potential of a terrestrial vascular plant, Portulaca Oleracea L. as a biomonitoring tool of fresh water aquatic environments.

    PubMed

    Anandi, S; Thangavel, P; Subburam, V

    2002-08-01

    In the present study, the influence of aluminium on the regeneration potential of leaf and stem-cuttings of Portulaca oleracea was studied in order to identify a terrestrial plant species as an alternate biomonitoring tool of fresh water environment. The leaves and stem cuttings of the terrestrial plant, P. oleracea grew well in the distilled water producing adventitious and lateral roots. The aluminium treated leaves and stem cuttings showed a reduction in the growth of the adventitious and lateral roots and increase in the decay of leaves and stem cuttings with increasing aluminium concentration. The tolerance index calculated for the leaves and the stem cuttings showed that the leaves were more sensitive than stems to aluminium. Since all the parameters studied showed concentration dependent changes, the terrestrial plant, P oleracea can be considered a suitable 'biomonitoring tool' of fresh water environment besides its usage in the Al toxicity testing. PMID:12197638

  14. [Some aspects of underground organs of spotleaf orchis growth and phenolic compound accumulation at the generative stage of ontogenesis].

    PubMed

    Marakaev, O A; Tselebrovskiĭ, M V; Nikolaeva, T N; Zagoskina, N V

    2013-01-01

    For the first time, the time courses of the growth and accumulation of phenolic compounds (PHCs) in different parts of the organs of the underground organs of the tuberoid orchid Dactilorhyza maculata during vegetation and the winter dormancy period were studied. It was shown that the replacement of old tuberoids and adventitious roots by new ones is accompanied by active growth, photosynthate redistribution, and PHC accumulation. It was found that the tuberoid performing the function of a storage organ possessed a low PHC content, which increased at the end of vegetation and was stably high during the winter dormancy period; in spring the content decreased, most likely as a result of PHC catabolism and utilization for growth processes. It was proposed that the higher PHC content in adventitious roots and tuberoid terminal parts is determined by mycotrophy of the organs.

  15. [Genetic regulation of plant shoot stem cells].

    PubMed

    Al'bert, E V; Ezhova, T A

    2013-02-01

    This article describes the main features of plant stem cells and summarizes the results of studies of the genetic control of stem cell maintenance in the apical meristem of the shoot. It is demonstrated that the WUS-CLV gene system plays a key role in the maintenance of shoot apical stem cells and the formation of adventitious buds and somatic embryos. Unconventional concepts of plant stem cells are considered.

  16. Pollen- and Seed-Mediated Transgene Flow in Commercial Cotton Seed Production Fields

    PubMed Central

    Heuberger, Shannon; Ellers-Kirk, Christa; Tabashnik, Bruce E.; Carrière, Yves

    2010-01-01

    Background Characterizing the spatial patterns of gene flow from transgenic crops is challenging, making it difficult to design containment strategies for markets that regulate the adventitious presence of transgenes. Insecticidal Bacillus thuringiensis (Bt) cotton is planted on millions of hectares annually and is a potential source of transgene flow. Methodology/Principal Findings Here we monitored 15 non-Bt cotton (Gossypium hirsutum, L.) seed production fields (some transgenic for herbicide resistance, some not) for gene flow of the Bt cotton cry1Ac transgene. We investigated seed-mediated gene flow, which yields adventitious Bt cotton plants, and pollen-mediated gene flow, which generates outcrossed seeds. A spatially-explicit statistical analysis was used to quantify the effects of nearby Bt and non-Bt cotton fields at various spatial scales, along with the effects of pollinator abundance and adventitious Bt plants in fields, on pollen-mediated gene flow. Adventitious Bt cotton plants, resulting from seed bags and planting error, comprised over 15% of plants sampled from the edges of three seed production fields. In contrast, pollen-mediated gene flow affected less than 1% of the seed sampled from field edges. Variation in outcrossing was better explained by the area of Bt cotton fields within 750 m of the seed production fields than by the area of Bt cotton within larger or smaller spatial scales. Variation in outcrossing was also positively associated with the abundance of honey bees. Conclusions/Significance A comparison of statistical methods showed that our spatially-explicit analysis was more powerful for understanding the effects of surrounding fields than customary models based on distance. Given the low rates of pollen-mediated gene flow observed in this study, we conclude that careful planting and screening of seeds could be more important than field spacing for limiting gene flow. PMID:21152426

  17. First megafossil evidence of Cyatheaceous tree fern from the Indian Cenozoic

    NASA Astrophysics Data System (ADS)

    Bera, Subir; Gupta, Sudha; Khan, Mahasin Ali; De, Aniruddha; Mukhopadhyay, Radhanath

    2014-08-01

    A part of the compressed tree fern axis with leaf and adventitious root scars in unusual arrangement from Plio-Pleistocene sediments of Arunachal Pradesh, India is described as Cyathea siwalika sp. nov. This record suggests that Cyathea was an important component of tropical evergreen forest in the area during the Plio-Pleistocene and this group subsequently declined in importance within the local vegetation.

  18. Early grass seedling growth stage improves explanation of future stand success

    SciTech Connect

    Ries, R.E.

    1999-07-01

    Predicting future grass stand success shortly after seeding improves the author's understanding of the mechanism of seeding success and makes possible timely decisions on the potential productivity of new grass stands. A 3-year field study with 5 species and 11 seeding dates per year was conducted to evaluate grass development and success when grass was direct seeded into wheat stubble with a double disk come seeder with depth bands and packer wheels. The number of grass seedlings/m{sup 2}, the number of adventitious roots, Haun stage, leaf length, leaf area, and number of tillers were measured 45 days after emergence. Earlier results showed that grass stand success can be reasonably predicted from the relationship of seedlings/m{sup 2} at 45 days after emergence with grass stems or plants/m{sup 2} 2 years after seeding. However, the simple coefficient of determination (r{sup 2}) for these relationships was quite low. further multiple regression analysis has shown that the R{sup 2} values can be significantly improved by adding the appropriate seedling growth stage to the number of seedlings/m{sup 2} at 45 days. For smooth bromegrass, the r{sup 2} was improved to 0.75 with the addition of adventitious roots, leaf area, and leaf length. The R{sup 2} was increased to 0.58 for crested wheatgrass and improved to 0.38 for western wheatgrass with the addition of number of adventitious roots. The R{sup 2} for blue grama was increased to 0.65 with the addition of number of adventitious roots. These data show that the explanation of the number of stems or plants/m{sup 2} estimated 2 years into the future is improved by knowing something about the developmental stage of the seedlings/m{sup 2} at 45 days after emergence.

  19. Cross-fertilization between genetically modified and non-genetically modified maize crops in Uruguay.

    PubMed

    Galeano, Pablo; Debat, Claudio Martínez; Ruibal, Fabiana; Fraguas, Laura Franco; Galván, Guillermo A

    2010-01-01

    The cultivation of genetically modified (GM) Bt maize (Zea mays L.) events MON810 and Bt11 is permitted in Uruguay. Local regulations specify that 10% of the crop should be a non-GM cultivar as refuge area for biodiversity, and the distance from other non-GM maize crops should be more than 250 m in order to avoid cross-pollination. However, the degree of cross-fertilization between maize crops in Uruguay is unknown. The level of adventitious presence of GM material in non-GM crops is a relevant issue for organic farming, in situ conservation of genetic resources and seed production. In the research reported here, the occurrence and frequency of cross-fertilization between commercial GM and non-GM maize crops in Uruguay was assessed. The methodology comprised field sampling and detection using DAS-ELISA and PCR. Five field-pair cases where GM maize crops were grown near non-GM maize crops were identified. These cases had the potential to cross-fertilize considering the distance between crops and the similarity of the sowing dates. Adventitious presence of GM material in the offspring of non-GM crops was found in three of the five cases. Adventitious presence of event MON810 or Bt11 in non-GM maize, which were distinguished using specific primers, matched the events in the putative sources of transgenic pollen. Percentages of transgenic seedlings in the offspring of the non-GM crops were estimated as 0.56%, 0.83% and 0.13% for three sampling sites with distances of respectively 40, 100 and 330 m from the GM crops. This is a first indication that adventitious presence of transgenes in non-GM maize crops will occur in Uruguay if isolation by distance and/or time is not provided. These findings contribute to the evaluation of the applicability of the "regulated coexistence policy" in Uruguay.

  20. The development of Archaeopteris: new evolutionary characters from the structural analysis of an Early Famennian trunk from southeast Morocco.

    PubMed

    Meyer-Berthaud, B; Scheckler, S E; Bousquet, J L

    2000-04-01

    A 5 m long trunk of a young Archaeopteris/Callixylon erianum tree from the Late Devonian of Morocco shows new branching patterns for early lignophytes. This progymnosperm tree produces a helical pattern of traces that we infer belonged to reduced, short-lived, primary (apical) branches (type A) as well as two types of adventitious traces (types B and H). We infer that type-B traces supplied branches that initiate close to the site of attachment on the trunk of some, but not all type-A branches in an irregular but nonrandom pattern. Unlike ephemeral type-A branches, those of type B persist and become long-lived, potentially permanent units of the architecture of Archaeopteris trees. Type-H adventitious traces are also short-lived and occur singly or in serial groups, but differ from traces of either type A or B branches by lacking differentiation into a readily identifiable organ category. We interpret type-H traces as supplying latent primordia that could develop into either adventitious roots or shoots depending on extrinsic factors. Our new data suggest that Archaeopteris had a wide range of branch primordium amplitude. Type-B branches compare with axillary lateral branch buds of some Early Carboniferous spermatophytes (Calamopitys) and are a major developmental departure from the strictly apical, pseudomonopodial shoot branching of older aneurophyte progymnosperms. Type-H traces suggest that Archaeopteris trees had some potential for formation of adventitious roots or shoots in response to environmental factors, such as partial burial by overbank sedimentation. Collectively, these novel methods of tree branching may partly explain the extraordinary success and worldwide dominance of Archaeopteris forests on fluvially dominated, Late Devonian floodplains.

  1. Process-Induced Carbon and Sub-Layer in SiC/BN/SiC Composites: Characterization and Consequences

    NASA Technical Reports Server (NTRS)

    Ogbuji, L. U. J. T; Wheeler, D. R.; McCue, T. R.

    2001-01-01

    Following our detection of films of elemental carbon in the Hi-Nicalon TM/BN/SiC composite and its deleterious effect on oxidative durability, we have examined other SiC/BN/SiC systems. The problem is pervasive, and significant residues of free carbon are confirmed in Sylramic /BN/SiC materials. Effective techniques for routine detection and characterization of adventitious carbon in SiC/BN/SiC composites are discussed.

  2. Collagen Matrix Remodeling in Stented Pulmonary Arteries after Transapical Heart Valve Replacement.

    PubMed

    Ghazanfari, Samaneh; Driessen-Mol, Anita; Hoerstrup, Simon P; Baaijens, Frank P T; Bouten, Carlijn V C

    2016-01-01

    The use of valved stents for minimally invasive replacement of semilunar heart valves is expected to change the extracellular matrix and mechanical function of the native artery and may thus impair long-term functionality of the implant. Here we investigate the impact of the stent on matrix remodeling of the pulmonary artery in a sheep model, focusing on matrix composition and collagen (re)orientation of the host tissue. Ovine native pulmonary arteries were harvested 8 (n = 2), 16 (n = 4) and 24 (n = 2) weeks after transapical implantation of self-expandable stented heart valves. Second harmonic generation (SHG) microscopy was used to assess the collagen (re)orientation of fresh tissue samples. The collagen and elastin content was quantified using biochemical assays. SHG microscopy revealed regional differences in collagen organization in all explants. In the adventitial layer of the arterial wall far distal to the stent (considered as the control tissue), we observed wavy collagen fibers oriented in the circumferential direction. These circumferential fibers were more straightened in the adventitial layer located behind the stent. On the luminal side of the wall behind the stent, collagen fibers were aligned along the stent struts and randomly oriented between the struts. Immediately distal to the stent, however, fibers on both the luminal and the adventitial side of the wall were oriented in the axial direction, demonstrating the stent impact on the collagen structure of surrounding arterial tissues. Collagen orientation patterns did not change with implantation time, and biochemical analyses showed no changes in the trend of collagen and elastin content with implantation time or location of the vascular wall. We hypothesize that the collagen fibers on the adventitial side of the arterial wall and behind the stent straighten in response to the arterial stretch caused by oversizing of the stent. However, the collagen organization on the luminal side suggests that

  3. Theoretical effects of UTB urea transporters in the renal medullary microcirculation.

    PubMed

    Zhang, Wensheng; Edwards, Aurélie

    2003-10-01

    A mathematical model of transport in the renal medullary microcirculation was used to investigate the role of the UTB urea transporter expressed in descending vasa recta (DVR) endothelia and red blood cell (RBC) membranes. Our simulations suggest that UTB raises RBC and plasma and interstitial urea concentrations by facilitating radial diffusion of the solute and therefore serves to increase the contribution of urea to the corticomedullary osmolality gradient, assuming no secondary effects on tubular transport. However, by lowering transmural urea concentration gradients, UTB reduces water efflux from DVR through aquaporin-1 (AQP1) water channels, thereby decreasing plasma sodium concentration. The net result of these competing effects on the osmolality gradient depends on the fraction of filtered urea that is reabsorbed by vasa recta. We also found that the contribution of UTB to water transport across DVR and RBCs is negligible, even in the absence of AQP1. Our model predicts that UTB plays a significant role, however, in reducing the shrinking and swelling of RBCs as blood flows along the medulla. PMID:12824077

  4. Germ cell differentiation and proliferation in the developing testis of the South American plains viscacha, Lagostomus maximus (Mammalia, Rodentia).

    PubMed

    Gonzalez, C R; Muscarsel Isla, M L; Fraunhoffer, N A; Leopardo, N P; Vitullo, A D

    2012-08-01

    Cell proliferation and cell death are essential processes in the physiology of the developing testis that strongly influence the normal adult spermatogenesis. We analysed in this study the morphometry, the expression of the proliferation cell nuclear antigen (PCNA), cell pluripotency marker OCT-4, germ cell marker VASA and apoptosis in the developing testes of Lagostomus maximus, a rodent in which female germ line develops through abolished apoptosis and unrestricted proliferation. Morphometry revealed an increment in the size of the seminiferous cords with increasing developmental age, arising from a significant increase of PCNA-positive germ cells and a stable proportion of PCNA-positive Sertoli cells. VASA showed a widespread cytoplasmic distribution in a great proportion of proliferating gonocytes that increased significantly at late development. In the somatic compartment, Leydig cells increased at mid-development, whereas peritubular cells showed a stable rate of proliferation. In contrast to other mammals, OCT-4 positive gonocytes increased throughout development reaching 90% of germ cells in late-developing testis, associated with a conspicuous increase in circulating FSH from mid- to late-gestation. TUNEL analysis was remarkable negative, and only a few positive cells were detected in the somatic compartment. These results show that the South American plains viscacha displays a distinctive pattern of testis development characterized by a sustained proliferation of germ cells throughout development, with no signs of apoptosis cell demise, in a peculiar endocrine in utero ambiance that seems to promote the increase of spermatogonial number as a primary direct effect of FSH.

  5. Germ cell degeneration in high-temperature treated pufferfish, Takifugu rubripes.

    PubMed

    Lee, K H; Yamaguchi, A; Rashid, H; Kadomura, K; Yasumoto, S; Matsuyama, M

    2009-01-01

    Exogenous factors such as temperature, social behavior, and salinity play a crucial role during the critical sensitive period of sex differentiation in many vertebrates. In fishes, amphibians, and reptiles temperature treatment is known to induce all-male (or female) individuals, and genes related to sex differentiation have been studied. The Japanese pufferfish, Takifugu rubripes, possesses the most compact genome among vertebrates and has immense potential for studies focusing on comparative genome analysis. In this study, we describe gonadal morphology and vasa (germ cell marker) and dmrt1 (Sertoli cell marker) expression on a molecular level in relation to the development of temperature-treated pufferfish. To investigate the relationship between temperature and gonadal development, pufferfish were exposed to high-temperature conditions (32 degrees C) during early gonadal development. Morphological observations showed that this high-temperature treatment did not influence sexual differentiation as determined by ovarian cavity characteristics; however, high-temperature treatment induces gonadal degeneration that is devoid of germ cells. RT-PCR results revealed no vasa expression within germ cell-degenerated gonads. In situ hybridization results showed that dmrt1 was expressed in somatic cells of germ cell-degenerated ovaries. These results suggest that high-temperature treatment during early gonadal development induces germ cell degeneration and masculinization of ovarian somatic cells in pufferfish.

  6. THY1 as a reliable marker for enrichment of undifferentiated spermatogonia in the goat.

    PubMed

    Abbasi, Hassan; Tahmoorespur, Mojtaba; Hosseini, Sayyed Morteza; Nasiri, Zahra; Bahadorani, Mehrnoosh; Hajian, Mahdi; Nasiri, Mohammad Reza; Nasr-Esfahani, Mohammad Hossein

    2013-11-01

    Spermatogonial stem cells are unique cells of testes that can restore fertility upon transplantation into recipient testes. However, use of suitable markers for enrichment of these cells have important potential application. THY1, is an established conserved marker of spermatogonial stem cells in bovine, rodents, and primates, but there is no information available in goats. After three rounds of enzymatic digestion of prepubertal goat testicular tissues, undifferentiated spermatogonia positive for THY1 were isolated by magnetic-activated cell sorting and were used for immunocytochemistry, real-time polymerase chain reaction analysis for gene expression, protein expression, and transplantation into recipient mice. Immunocytochemical analyses showed that significantly higher percentage of THY1(+) cells were positive for PLZF and VASA when compared with unselected population. This result for PLZF was further confirmed at the protein level. Real-time polymerase chain reaction analysis revealed that expression of THY1, PLZF, VASA, BCL6B, and UCHL1 as SCCs characteristic genes in THY1(+) cells was significantly higher than in the initial population. Finally, transplantation of PKH26-labeled cells revealed that THY1(+) cells had higher capacity for colony formation when compared with unselected cells. In conclusion, the results provide indications that THY1 surface marker can be reliably used for enrichment of undifferentiated spermatogonial in the goats.

  7. Protective effect of vitamin E on methyl methanesulfonate-induced teratozoospermia in adult Sprague-Dawley rats.

    PubMed

    Tang, Zhian; Ding, Weiliang; Wang, Lun; Jiang, Wenchu; Zhang, Quanxiang; Chen, Hong; Zou, Hongnan; Dong, Yongkang; Shao, Jianwei; Ma, Tieliang

    2015-09-01

    The protective effect of vitamin E (VE, α-tocopherol) on methyl methanesulfonate (MMS)-induced teratozoospermia was investigated in adult rats. Rats (n=6 per group) were divided into three groups: i) Control group, treated with distilled water from days 1 to 5; ii) the MMS group, treated with MMS at a dose of 40 mg·kg(-1) from days 1‑5; or iii) the VE+MMS group, treated with MMS at a dose of 40 mg·kg(-1) from days 1‑5, followed by VE at a dose of 150 mg·kg(-1) from day 6 for 6 weeks. Sperm count, motility and morphology were examined following treatment with VE. The serum testosterone level and antioxidant enzyme activity were measured, and the localization of Vasa, promyelocytic leukemia zinc finger protein (Plzf) and synaptonemal complex protein 3 (Scp3) were also examined. MMS treatment decreased sperm count and motility, and the levels of immunoreactive serum testosterone and endogenous antioxidants. In addition, MMS increased the percentage of abnormal sperm and the levels of free radicals. After MMS and VE treatment, sperm count and motility were significantly higher in rats from the VE+MMS group than in the MMS group. In addition, the serum testosterone concentration, as well as the levels of Vasa and free radicals and the percentage of abnormal sperm, decreased. The results indicated that VE has protective effects against MMS-induced teratozoospermia in adult rats.

  8. [Vasculitic Peripheral Neuropathies: Clinical Features and Diagnostic Laboratory Tests].

    PubMed

    Ogata, Katsuhisa

    2016-03-01

    Vasculitic peripheral neuropathy (VPN) occurs due to ischemic changes of peripheral nerves, resulting from a deficit of vascular blood supply due to damaged vasa nervorum leading to vasculitis. VPN usually manifests as sensorimotor or sensory disturbances accompanied by pain, presenting as a type of multiple mononeuropathy, with a scattered distribution in distal limbs. VPN may also present as a mononeuropathy, distal symmetric polyneuropathy, plexopathy, or radiculopathy. The rapidity of VPN is variable, ranging from days to months, with symptoms occasionally changing with the appearance of new lesions. Careful history taking and neurological examination provides an exact diagnosis. The most common cause of VPN is primary vasculitis predominantly affecting small vessels, including vasa nervorum, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, and polyarteritis nodosa. Similar vasculitic processes can also result from a systemic collagen disorder or secondary vasculitis. Electrophysiological studies and pathological investigation of biopsied peripheral nerves and muscles are important for diagnosis of vasculitis. Serological tests, including ANCA, are useful for diagnosis of vasculitis. Accurate neurological examinations are essential for diagnosis and evaluation of clinical course.

  9. Catheter fragmentation and local lysis in two lung transplant patients with pulmonary embolism.

    PubMed

    Suhling, H; Westerkamp, V; Dinh, Q T; Greer, M; Kempf, T; Simon, A; Gottlieb, J; Welte, T; Schieffer, B

    2010-11-01

    In conjunction with the rising number of lung transplant operations in the past decade, an increased predisposition to venous thrombosis (VT), particularly within the first year posttransplantation has been observed. Previous studies have revealed that between 8.6% and 12% of patients develop VT, which can ultimately result in pulmonary emboli (PE).Transplanted lungs pose a much greater infarction risk due to their lack of collateral vascularisation, relying entirely on the vasa publica--the pulmonary artery--in the absence of vasa privata. Such losses in viable lung parenchyma are always serious, but carry still greater risks for single-lung transplant recipients, an early diagnosis and treatment remain critical. Here we report on two cases of PE after lung transplantation, both of whom were managed with catheter fragmentation and local thrombolysis. In our opinion, this approach represents a viable treatment for symptomatic PE in lung transplant recipients. The benefits and risks of the alternative treatment options in these special cases will be reviewed and the definitive therapy was described. In the patients treated, catheter fragmentation with localized thrombolysis resulted in short term improvements in graft function, but could not prevent later lung infarction in one case.

  10. Modulation of gurken translation by insulin and TOR signaling in Drosophila

    PubMed Central

    Ferguson, Scott B.; Blundon, Malachi A.; Klovstad, Martha S.; Schüpbach, Trudi

    2012-01-01

    Localized Gurken (Grk) translation specifies the anterior–posterior and dorsal–ventral axes of the developing Drosophila oocyte; spindle-class females lay ventralized eggs resulting from inefficient grk translation. This phenotype is thought to result from inhibition of the Vasa RNA helicase. In a screen for modifiers of the eggshell phenotype in spn-B flies, we identified a mutation in the lnk gene. We show that lnk mutations restore Grk expression but do not suppress the persistence of double-strand breaks nor other spn-B phenotypes. This suppression does not affect Egfr directly, but rather overcomes the translational block of grk messages seen in spindle mutants. Lnk was recently identified as a component of the insulin/insulin-like growth factor signaling (IIS) and TOR pathway. Interestingly, direct inhibition of TOR with rapamycin in spn-B or vas mutant mothers can also suppress the ventralized eggshell phenotype. When dietary protein is inadequate, reduced IIS–TOR activity inhibits cap-dependent translation by promoting the activity of the translation inhibitor eIF4E-binding protein (4EBP). We hypothesize that reduced TOR activity promotes grk translation independent of the canonical Vasa- and cap-dependent mechanism. This model might explain how flies can maintain the translation of developmentally important transcripts during periods of nutrient limitation when bulk cap-dependent translation is repressed. PMID:22328499

  11. [Vasculitic Peripheral Neuropathies: Clinical Features and Diagnostic Laboratory Tests].

    PubMed

    Ogata, Katsuhisa

    2016-03-01

    Vasculitic peripheral neuropathy (VPN) occurs due to ischemic changes of peripheral nerves, resulting from a deficit of vascular blood supply due to damaged vasa nervorum leading to vasculitis. VPN usually manifests as sensorimotor or sensory disturbances accompanied by pain, presenting as a type of multiple mononeuropathy, with a scattered distribution in distal limbs. VPN may also present as a mononeuropathy, distal symmetric polyneuropathy, plexopathy, or radiculopathy. The rapidity of VPN is variable, ranging from days to months, with symptoms occasionally changing with the appearance of new lesions. Careful history taking and neurological examination provides an exact diagnosis. The most common cause of VPN is primary vasculitis predominantly affecting small vessels, including vasa nervorum, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, and polyarteritis nodosa. Similar vasculitic processes can also result from a systemic collagen disorder or secondary vasculitis. Electrophysiological studies and pathological investigation of biopsied peripheral nerves and muscles are important for diagnosis of vasculitis. Serological tests, including ANCA, are useful for diagnosis of vasculitis. Accurate neurological examinations are essential for diagnosis and evaluation of clinical course. PMID:27001769

  12. Cladistic analysis of Subfamily Bruchomyiinae (Diptera: Psychodidae).

    PubMed

    Wagner, Rüdiger; Stuckenberg, Brian

    2016-01-01

    Subfamily Bruchomyiinae is comprised of 60 species and has been referred to as the most primitive within the Psychodidae. The assumed sister-group relationship with Phlebotominae is based on ecological constraints of their environment. A cladistics analysis based on 29 characters and 52 species revealed the distinction of an Old World clade characterized by males with elongate, narrow vasa deferentia, and a New World clade with males having shorter and basally widened vasa deferentia. The Old World clade consists of the genera Nemopalpus Macquart (9 species), and Eutonnoiria Alexander (1 species). The New World clade includes Bruchomyia Alexander (10 species), Boreofairchildia genus nov. (13 species), Laurenceomyia genus nov. (5 species), and Notofairchildia genus nov. (15 species). Parsimony and Bayesian analyses resulted in trees that generally support this generic classification; however, with some species groups less resolved. Diagnostic features for genera are provided. In contrast to the other New World genera, Notofairchildia is paraphyletic with the provisional inclusion of at least the Australasian taxa. PMID:27394447

  13. Dual Angiogenic and Neurotrophic Effects of Bone Marrow–Derived Endothelial Progenitor Cells on Diabetic Neuropathy

    PubMed Central

    Jeong, Jin-Ok; Kim, Mee-Ohk; Kim, Hyongbum; Lee, Min-Young; Kim, Sung-Whan; Ii, Masaaki; Lee, Jung-uek; Lee, Jiyoon; Choi, Yong Jin; Cho, Hyun-Jai; Lee, Namho; Silver, Marcy; Wecker, Andrea; Kim, Dong-Wook; Yoon, Young-sup

    2009-01-01

    Background Endothelial progenitor cells (EPCs) are known to promote neovascularization in ischemic diseases. Recent evidence suggested that diabetic neuropathy is causally related to impaired angiogenesis and deficient growth factors. Accordingly, we investigated whether diabetic neuropathy could be reversed by local transplantation of EPCs. Methods and Results We found that motor and sensory nerve conduction velocities, blood flow, and capillary density were reduced in sciatic nerves of streptozotocin-induced diabetic mice but recovered to normal levels after hind-limb injection of bone marrow–derived EPCs. Injected EPCs were preferentially and durably engrafted in the sciatic nerves. A portion of engrafted EPCs were uniquely localized in close proximity to vasa nervorum, and a smaller portion of these EPCs were colocalized with endothelial cells. Multiple angiogenic and neurotrophic factors were significantly increased in the EPC-injected nerves. These dual angiogenic and neurotrophic effects of EPCs were confirmed by higher proliferation of Schwann cells and endothelial cells cultured in EPC-conditioned media. Conclusions We demonstrate for the first time that bone marrow-derived EPCs could reverse various manifestations of diabetic neuropathy. These therapeutic effects were mediated by direct augmentation of neovascularization in peripheral nerves through long-term and preferential engraftment of EPCs in nerves and particularly vasa nervorum and their paracrine effects. These findings suggest that EPC transplantation could represent an innovative therapeutic option for treating diabetic neuropathy. PMID:19171856

  14. Maintenance of somatic tissue regeneration with age in short- and long-lived species of sea urchins.

    PubMed

    Bodnar, Andrea G; Coffman, James A

    2016-08-01

    Aging in many animals is characterized by a failure to maintain tissue homeostasis and the loss of regenerative capacity. In this study, the ability to maintain tissue homeostasis and regenerative potential was investigated in sea urchins, a novel model to study longevity and negligible senescence. Sea urchins grow indeterminately, regenerate damaged appendages and reproduce throughout their lifespan and yet different species are reported to have very different life expectancies (ranging from 4 to more than 100 years). Quantitative analyses of cell proliferation and apoptosis indicated a low level of cell turnover in tissues of young and old sea urchins of species with different lifespans (Lytechinus variegatus, Strongylocentrotus purpuratus and Mesocentrotus franciscanus). The ability to regenerate damaged tissue was maintained with age as assessed by the regrowth of amputated spines and tube feet (motor and sensory appendages). Expression of genes involved in cell proliferation (pcna), telomere maintenance (tert) and multipotency (seawi and vasa) was maintained with age in somatic tissues. Immunolocalization of the Vasa protein to areas of the tube feet, spines, radial nerve, esophagus and a sub-population of circulating coelomocytes suggests the presence of multipotent cells that may play a role in normal tissue homeostasis and the regenerative potential of external appendages. The results indicate that regenerative potential was maintained with age regardless of lifespan, contrary to the expectation that shorter lived species would invest less in maintenance and repair. PMID:27095483

  15. Structure of the eastern Seattle fault zone, Washington state: New insights from seismic reflection data

    USGS Publications Warehouse

    Liberty, L.M.; Pratt, T.L.

    2008-01-01

    We identify and characterize the active Seattle fault zone (SFZ) east of Lake Washington with newly acquired seismic reflection data. Our results focus on structures observed in the upper 1 km below the cities of Bellevue, Sammamish, Newcastle, and Fall City, Washington. The SFZ appears as a broad zone of faulting and folding at the southern boundary of the Seattle basin and north edge of the Seattle uplift. We interpret the Seattle fault as a thrust fault that accommodates north-south shortening by forming a fault-propagation fold with a forelimb breakthrough. The blind tip of the main fault forms a synclinal growth fold (deformation front) that extends at least 8 km east of Vasa Park (west side of Lake Sammamish) and defines the south edge of the Seattle basin. South of the deformation front is the forelimb break-through fault, which was exposed in a trench at Vasa Park. The Newcastle Hills anticline, a broad anticline forming the north part of the Seattle uplift east of Lake Washington, is interpreted to lie between the main blind strand of the Seattle fault and a backthrust. Our profiles, on the northern limb of this anticline, consistently image north-dipping strata. A structural model for the SFZ east of Lake Washington is consistent with about 8 km of slip on the upper part of the Seattle fault, but the amount of motion is only loosely constrained.

  16. Cladistic analysis of Subfamily Bruchomyiinae (Diptera: Psychodidae).

    PubMed

    Wagner, Rüdiger; Stuckenberg, Brian

    2016-01-01

    Subfamily Bruchomyiinae is comprised of 60 species and has been referred to as the most primitive within the Psychodidae. The assumed sister-group relationship with Phlebotominae is based on ecological constraints of their environment. A cladistics analysis based on 29 characters and 52 species revealed the distinction of an Old World clade characterized by males with elongate, narrow vasa deferentia, and a New World clade with males having shorter and basally widened vasa deferentia. The Old World clade consists of the genera Nemopalpus Macquart (9 species), and Eutonnoiria Alexander (1 species). The New World clade includes Bruchomyia Alexander (10 species), Boreofairchildia genus nov. (13 species), Laurenceomyia genus nov. (5 species), and Notofairchildia genus nov. (15 species). Parsimony and Bayesian analyses resulted in trees that generally support this generic classification; however, with some species groups less resolved. Diagnostic features for genera are provided. In contrast to the other New World genera, Notofairchildia is paraphyletic with the provisional inclusion of at least the Australasian taxa.

  17. Conservation and expression of PIWI-interacting RNA pathway genes in male and female adult gonad of amniotes.

    PubMed

    Lim, Shu Ly; Tsend-Ayush, Enkhjargal; Kortschak, R Daniel; Jacob, Reuben; Ricciardelli, Carmela; Oehler, Martin K; Grützner, Frank

    2013-12-01

    The PIWI-interacting RNA (piRNA) pathway is essential for germline development and transposable element repression. Key elements of this pathway are members of the piRNA-binding PIWI/Argonaute protein family and associated factors (e.g., VASA, MAELSTROM, and TUDOR domain proteins). PIWI-interacting RNAs have been identified in mouse testis and oocytes, but information about the expression of the different piRNA pathway genes, in particular in the mammalian ovary, remains incomplete. We investigated the evolution and expression of piRNA pathway genes in gonads of amniote species (chicken, platypus, and mouse). Database searches confirm a high level of conservation and revealed lineage-specific gain and loss of Piwi genes in vertebrates. Expression analysis in mammals shows that orthologs of Piwi-like (Piwil) genes, Mael (Maelstrom), Mvh (mouse vasa homolog), and Tdrd1 (Tudor domain-containing protein 1) are expressed in platypus adult testis. In contrast to mouse, Piwil4 is expressed in platypus and human adult testis. We found evidence for Mael and Piwil2 expression in mouse Sertoli cells. Importantly, we show mRNA expression of Piwil2, Piwil4, and Mael in oocytes and supporting cells of human, mouse, and platypus ovary. We found no Piwil1 expression in mouse and chicken ovary. The conservation of gene expression in somatic parts of the gonad and germ cells of species that diverged over 800 million yr ago indicates an important role in adult male and female gonad. PMID:24108303

  18. The DEAD Box RNA helicase VBH-1 is a new player in the stress response in C. elegans.

    PubMed

    Paz-Gómez, Daniel; Villanueva-Chimal, Emmanuel; Navarro, Rosa E

    2014-01-01

    For several years, DEAD box RNA helicase Vasa (DDX4) has been used as a bona fide germline marker in different organisms. C. elegans VBH-1 is a close homolog of the Vasa protein, which plays an important role in gametogenesis, germ cell survival and embryonic development. Here, we show that VBH-1 protects nematodes from heat shock and oxidative stress. Using the germline-defective mutant glp-4(bn2) we found that a potential somatic expression of vbh-1 might be important for stress survival. We also show that the VBH-1 paralog LAF-1 is important for stress survival, although this protein is not redundant with its counterpart. Furthermore, we observed that the mRNAs of the heat shock proteins hsp-1 and sip-1 are downregulated when vbh-1 or laf-1 are silenced. Previously, we reported that in C. elegans, VBH-1 was primarily expressed in P granules of germ cells and in the cytoplasm of all blastomeres. Here we show that during stress, VBH-1 co-localizes with CGH-1 in large aggregates in the gonad core and oocytes; however, VBH-1 aggregates do not overlap with CGH-1 foci in early embryos under the same conditions. These data demonstrate that, in addition to the previously described role for this protein in the germline, VBH-1 plays an important role during the stress response in C. elegans through the potential direct or indirect regulation of stress response mRNAs.

  19. Evidence against a germ plasm in the milkweed bug Oncopeltus fasciatus, a hemimetabolous insect.

    PubMed

    Ewen-Campen, Ben; Jones, Tamsin E M; Extavour, Cassandra G

    2013-06-15

    Primordial germ cell (PGC) formation in holometabolous insects like Drosophila melanogaster relies on maternally synthesised germ cell determinants that are asymmetrically localised to the oocyte posterior cortex. Embryonic nuclei that inherit this "germ plasm" acquire PGC fate. In contrast, historical studies of basally branching insects (Hemimetabola) suggest that a maternal requirement for germ line genes in PGC specification may be a derived character confined principally to Holometabola. However, there have been remarkably few investigations of germ line gene expression and function in hemimetabolous insects. Here we characterise PGC formation in the milkweed bug Oncopeltus fasciatus, a member of the sister group to Holometabola, thus providing an important evolutionary comparison to members of this clade. We examine the transcript distribution of orthologues of 19 Drosophila germ cell and/or germ plasm marker genes, and show that none of them localise asymmetrically within Oncopeltus oocytes or early embryos. Using multiple molecular and cytological criteria, we provide evidence that PGCs form after cellularisation at the site of gastrulation. Functional studies of vasa and tudor reveal that these genes are not required for germ cell formation, but that vasa is required in adult males for spermatogenesis. Taken together, our results provide evidence that Oncopeltus germ cells may form in the absence of germ plasm, consistent with the hypothesis that germ plasm is a derived strategy of germ cell specification in insects. PMID:23789106

  20. The diversity of nanos expression in echinoderm embryos supports different mechanisms in germ cell specification.

    PubMed

    Fresques, Tara; Swartz, Steven Zachary; Juliano, Celina; Morino, Yoshiaki; Kikuchi, Mani; Akasaka, Koji; Wada, Hiroshi; Yajima, Mamiko; Wessel, Gary M

    2016-07-01

    Specification of the germ cell lineage is required for sexual reproduction in all animals. However, the timing and mechanisms of germ cell specification is remarkably diverse in animal development. Echinoderms, such as sea urchins and sea stars, are excellent model systems to study the molecular and cellular mechanisms that contribute to germ cell specification. In several echinoderm embryos tested, the germ cell factor Vasa accumulates broadly during early development and is restricted after gastrulation to cells that contribute to the germ cell lineage. In the sea urchin, however, the germ cell factor Vasa is restricted to a specific lineage by the 32-cell stage. We therefore hypothesized that the germ cell specification program in the sea urchin/Euechinoid lineage has evolved to an earlier developmental time point. To test this hypothesis we determined the expression pattern of a second germ cell factor, Nanos, in four out of five extant echinoderm clades. Here we find that Nanos mRNA does not accumulate until the blastula stage or later during the development of all other echinoderm embryos except those that belong to the Echinoid lineage. Instead, Nanos is expressed in a restricted domain at the 32-128 cell stage in Echinoid embryos. Our results support the model that the germ cell specification program underwent a heterochronic shift in the Echinoid lineage. A comparison of Echinoid and non-Echinoid germ cell specification mechanisms will contribute to our understanding of how these mechanisms have changed during animal evolution. PMID:27402572

  1. Differential Expression of Conserved Germ Line Markers and Delayed Segregation of Male and Female Primordial Germ Cells in a Hermaphrodite, the Leech Helobdella

    PubMed Central

    Cho, Sung-Jin; Vallès, Yvonne; Weisblat, David A.

    2014-01-01

    In sexually reproducing animals, primordial germ cells (PGCs) are often set aside early in embryogenesis, a strategy that minimizes the risk of genomic damage associated with replication and mitosis during the cell cycle. Here, we have used germ line markers (piwi, vasa, and nanos) and microinjected cell lineage tracers to show that PGC specification in the leech genus Helobdella follows a different scenario: in this hermaphrodite, the male and female PGCs segregate from somatic lineages only after more than 20 rounds of zygotic mitosis; the male and female PGCs share the same (mesodermal) cell lineage for 19 rounds of zygotic mitosis. Moreover, while all three markers are expressed in both male and female reproductive tissues of the adult, they are expressed differentially between the male and female PGCs of the developing embryo: piwi and vasa are expressed preferentially in female PGCs at a time when nanos is expressed preferentially in male PGCs. A priori, the delayed segregation of male and female PGCs from somatic tissues and from one another increases the probability of mutations affecting both male and female PGCs of a given individual. We speculate that this suite of features, combined with a capacity for self-fertilization, may contribute to the dramatically rearranged genome of Helobdella robusta relative to other animals. PMID:24217283

  2. Modulation of gurken translation by insulin and TOR signaling in Drosophila.

    PubMed

    Ferguson, Scott B; Blundon, Malachi A; Klovstad, Martha S; Schüpbach, Trudi

    2012-03-15

    Localized Gurken (Grk) translation specifies the anterior-posterior and dorsal-ventral axes of the developing Drosophila oocyte; spindle-class females lay ventralized eggs resulting from inefficient grk translation. This phenotype is thought to result from inhibition of the Vasa RNA helicase. In a screen for modifiers of the eggshell phenotype in spn-B flies, we identified a mutation in the lnk gene. We show that lnk mutations restore Grk expression but do not suppress the persistence of double-strand breaks nor other spn-B phenotypes. This suppression does not affect Egfr directly, but rather overcomes the translational block of grk messages seen in spindle mutants. Lnk was recently identified as a component of the insulin/insulin-like growth factor signaling (IIS) and TOR pathway. Interestingly, direct inhibition of TOR with rapamycin in spn-B or vas mutant mothers can also suppress the ventralized eggshell phenotype. When dietary protein is inadequate, reduced IIS-TOR activity inhibits cap-dependent translation by promoting the activity of the translation inhibitor eIF4E-binding protein (4EBP). We hypothesize that reduced TOR activity promotes grk translation independent of the canonical Vasa- and cap-dependent mechanism. This model might explain how flies can maintain the translation of developmentally important transcripts during periods of nutrient limitation when bulk cap-dependent translation is repressed.

  3. Optical coherence tomography vs. high-frequency ultrasound during noninvasive laser coagulation of the canine vas deferens

    NASA Astrophysics Data System (ADS)

    Cilip, Christopher M.; Allaf, Mohamad E.; Fried, Nathaniel M.

    2012-02-01

    A noninvasive approach to vasectomy may eliminate male fear of complications related to surgery and increase its acceptance. Noninvasive laser thermal occlusion of the canine vas deferens has recently been reported. In this study, optical coherence tomography (OCT) and high-frequency ultrasound (HFUS) are compared for monitoring laser thermal coagulation of the vas in an acute canine model. Bilateral noninvasive laser coagulation of the vas was performed in 6 dogs (n=12 vasa) using a Ytterbium fiber laser wavelength of 1075 nm, incident power of 9.0 W, pulse duration of 500 ms, pulse rate of 1 Hz, and 3-mm-diameter spot. Cryogen spray cooling was used to prevent skin burns during the procedure. An OCT system with endoscopic probe and a HFUS system with 20-MHz transducer were used to image the vas immediately before and after the procedure. Vasa were then excised and processed for gross and histologic analysis for comparison with OCT and HFUS images. OCT provided high-resolution, superficial imaging of the compressed vas within the vas ring clamp, while HFUS provided deeper imaging of the vas held manually in the scrotal fold. Both OCT and high HFUS are promising imaging modalities for real-time confirmation of vas occlusion during noninvasive laser vasectomy.

  4. Application of optical coherence tomography and high-frequency ultrasound imaging during noninvasive laser vasectomy

    NASA Astrophysics Data System (ADS)

    Cilip, Christopher M.; Allaf, Mohamad E.; Fried, Nathaniel M.

    2012-04-01

    A noninvasive approach to vasectomy may eliminate male fear of complications related to surgery and increase its acceptance. Noninvasive laser thermal occlusion of the canine vas deferens has recently been reported. Optical coherence tomography (OCT) and high-frequency ultrasound (HFUS) are compared for monitoring laser thermal coagulation of the vas in an acute canine model. Bilateral noninvasive laser coagulation of the vas was performed in six dogs (n=12 vasa) using a Ytterbium fiber laser wavelength of 1075 nm, incident power of 9.0 W, pulse duration of 500 ms, pulse rate of 1 Hz, and 3-mm-diameter spot. Cryogen spray cooling was used to prevent skin burns during the procedure. An OCT system with endoscopic probe and a HFUS system with 20-MHz transducer were used to image the vas immediately before and after the procedure. Vasa were then excised and processed for gross and histologic analysis for comparison with OCT and HFUS images. OCT provided high-resolution, superficial imaging of the compressed vas within the vas ring clamp, while HFUS provided deeper imaging of the vas held manually in the scrotal fold. Both OCT and high HFUS are promising imaging modalities for real-time confirmation of vas occlusion during noninvasive laser vasectomy.

  5. Histopathologic Alterations Associated with Global Gene Expression Due to Chronic Dietary TCDD Exposure in Juvenile Zebrafish

    PubMed Central

    Liu, Qing; Spitsbergen, Jan M.; Cariou, Ronan; Huang, Chun-Yuan; Jiang, Nan; Goetz, Giles; Hutz, Reinhold J.; Tonellato, Peter J.; Carvan, Michael J.

    2014-01-01

    The goal of this project was to investigate the effects and possible developmental disease implication of chronic dietary TCDD exposure on global gene expression anchored to histopathologic analysis in juvenile zebrafish by functional genomic, histopathologic and analytic chemistry methods. Specifically, juvenile zebrafish were fed Biodiet starter with TCDD added at 0, 0.1, 1, 10 and 100 ppb, and fish were sampled following 0, 7, 14, 28 and 42 d after initiation of the exposure. TCDD accumulated in a dose- and time-dependent manner and 100 ppb TCDD caused TCDD accumulation in female (15.49 ppb) and male (18.04 ppb) fish at 28 d post exposure. Dietary TCDD caused multiple lesions in liver, kidney, intestine and ovary of zebrafish and functional dysregulation such as depletion of glycogen in liver, retrobulbar edema, degeneration of nasal neurosensory epithelium, underdevelopment of intestine, and diminution in the fraction of ovarian follicles containing vitellogenic oocytes. Importantly, lesions in nasal epithelium and evidence of endocrine disruption based on alternatively spliced vasa transcripts are two novel and significant results of this study. Microarray gene expression analysis comparing vehicle control to dietary TCDD revealed dysregulated genes involved in pathways associated with cardiac necrosis/cell death, cardiac fibrosis, renal necrosis/cell death and liver necrosis/cell death. These baseline toxicological effects provide evidence for the potential mechanisms of developmental dysfunctions induced by TCDD and vasa as a biomarker for ovarian developmental disruption. PMID:24988445

  6. Initiation of shoot apical meristem in rice: characterization of four SHOOTLESS genes.

    PubMed

    Satoh, N; Hong, S K; Nishimura, A; Matsuoka, M; Kitano, H; Nagato, Y

    1999-08-01

    The regulatory mechanism of shoot apical meristem (SAM) initiation is an important subject in developmental plant biology. We characterized nine recessive mutations derived from four independent loci (SHL1-SHL4) causing the deletion of the SAM. Radicles were produced in these mutant embryos. Concomitant with the loss of SAM, two embryo-specific organs, coleoptile and epiblast, were lost, but the scutellum was formed normally. Therefore, differentiation of radicle and scutellum is regulated independently of SAM, but that of coleoptile and epiblast may depend on SAM. Regeneration experiments using adventitious shoots from the scutellum-derived calli showed that no adventitious shoots were regenerated in any shl mutant. However, small adventitious leaves were observed in both mutant and wild-type calli, but they soon became necrotic and showed no extensive growth. Thus, leaf primordia can initiate in the absence of SAM, but their extensive growth requires the SAM. An in situ hybridization experiment using a rice homeobox gene, OSH1, as a probe revealed that shl1 and shl2 modified the expression domain of OSH1, but normal expression of OSH1 was observed in shl3 and shl4 embryos. Accordingly, SHL1 and SHL2 function upstream of OSH1, and SHL3 and SHL4 downstream or independently of OSH1. These shl mutants are useful for elucidating the genetic program driving SAM initiation and for unraveling the interrelationships among various organs in grass embryos. PMID:10409508

  7. Topography and immunocytochemical characterization of nerve fibers in the leptomeningeal compartments of the rat. A light- and electron-microscopical study.

    PubMed

    Fricke, B; von Düring, M; Andres, K H

    1997-01-01

    The localization of peptidergic, catecholaminergic, and nitroxidergic nerve fibers in the ventral leptomeningeal connective tissue compartment was studied in whole-mount preparations and serial semithin and ultrathin sections. For immunocytochemistry, whole-mount preparations of the leptomeninges and ventral brain slices with the meninges were incubated as free-floating specimens with primary antibodies against protein gene product 9.5 (PGP 9.5), substance P (SP), calcitonin gene-related peptide (CGRP), dopamine beta-hydroxylase (DbetaH), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and nitric oxide synthase (NOS) using the avidin-biotin-peroxidase method. Based on the regional differences of the connective tissue organization, the leptomeninx is subdivided into the pial, trabecular, and adventitial leptomeninx. The antibody PGP 9.5 stains all unmyelinated nerve fibers in the leptomeninx. Although the highest density of nerve fibers occurs in the adventitial leptomeninx, nerve fibers, and terminals are additionally present in the trabecular and pial leptomeninx. DbetaH-, NPY-, VIP- and NOS-immunoreactive (IR) nerve fibers occur exclusively in the adventitial leptomeninx forming neuromuscular junctions. CGRP- and SP-IR nerve fibers are localized in all three leptomeningeal compartments where they terminate close to the subarachnoid space (type 1) or within the connective tissue (type 2). Due to their morphological and immunocytochemical characterization a possible chemo-, mechano- or nociceptive function is discussed in the context of pathophysiological aspects.

  8. Wheezes, crackles and rhonchi: simplifying description of lung sounds increases the agreement on their classification: a study of 12 physicians' classification of lung sounds from video recordings

    PubMed Central

    Melbye, Hasse; Garcia-Marcos, Luis; Brand, Paul; Everard, Mark; Priftis, Kostas; Pasterkamp, Hans

    2016-01-01

    Background The European Respiratory Society (ERS) lung sounds repository contains 20 audiovisual recordings of children and adults. The present study aimed at determining the interobserver variation in the classification of sounds into detailed and broader categories of crackles and wheezes. Methods Recordings from 10 children and 10 adults were classified into 10 predefined sounds by 12 observers, 6 paediatricians and 6 doctors for adult patients. Multirater kappa (Fleiss' κ) was calculated for each of the 10 adventitious sounds and for combined categories of sounds. Results The majority of observers agreed on the presence of at least one adventitious sound in 17 cases. Poor to fair agreement (κ<0.40) was usually found for the detailed descriptions of the adventitious sounds, whereas moderate to good agreement was reached for the combined categories of crackles (κ=0.62) and wheezes (κ=0.59). The paediatricians did not reach better agreement on the child cases than the family physicians and specialists in adult medicine. Conclusions Descriptions of auscultation findings in broader terms were more reliably shared between observers compared to more detailed descriptions. PMID:27158515

  9. [Microscopic anatomy of abnormal structure in root tuber of Pueraria lobata].

    PubMed

    Duan, Hai-yan; Cheng, Ming-en; Peng, Hua-sheng; Zhang, He-ting; Zhao, Yu-jiao

    2015-11-01

    Puerariae Lobatae Radix, also known as Gegen, is a root derived from Pueraria lobata. Based on field investigation and the developmental anatomy of root tuber, we have elucidated the relationship between the growth of root tuber and the anomalous structure. The results of analysis showed that the root system of P. lobata was developed from seed and adventitious root and there existed root tuber, adventitious root and conductive root according to morphology and function. The root tuber was developed from adventitious root, its secondary structure conformed to the secondary structure of dicotyledon's root. With the development of root, the secondary phloem of root tuber appeared abnormal vascular tissue, which was distributed like ring in the outside of secondary vascular tissue. The root tuber might have 4-6 concentric circular permutation abnormal vascular tissuelobate, and was formed by the internal development of abnormal vascular tissue. The xylem and phloem of abnormal vascular tissue were the main body of the root tuber. The results reveal the abnormal anatomical structure development of P. lobata, also provides the theoretical basis for reasonable harvest medicinal parts and promoting sustainable utilization of resources of P. lobata.

  10. Somatic embryogenesis and enhanced shoot organogenesis in Metabriggsia ovalifolia W. T. Wang

    PubMed Central

    Ouyang, Yao; Chen, Yulu; Lü, Jinfeng; Teixeira da Silva, Jaime A.; Zhang, Xinhua; Ma, Guohua

    2016-01-01

    An efficient protocol providing a dual regeneration pathway via direct shoot organogenesis and somatic embryogenesis for an endangered species, Metabriggsia ovalifolia W. T. Wang, was established from leaf explants. When applied at 2.5 μM, the cytokinins 6-benzyladenine (BA) or thidiazuron (TDZ) and the auxins indole-3-butyric acid (IBA), α-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) could induce shoots when on basal Murashige and Skoog (MS) medium. BA and TDZ could induce more adventitious shoots (19.1 and 31.2/explant, respectively) than NAA (4.6/explant), IBA (5.7/explant) or IAA (6.4/explant). BA and TDZ at 5–10 μM could induce both shoots and somatic embryos. A higher concentration of TDZ (25 μM) induced only somatic embryos (39.8/explant). The same concentration of BA induced both adventitious shoots (23.6/explant) and somatic embryos (9.7/explant). Thus, somatic embryogenesis in this plant needs a high cytokinin concentration (BA; TDZ), as evidenced by histology. Somatic embryos germinated easily when left on the same media, but formed adventitious roots in two weeks on MS supplemented with 0.5 μM NAA, 0.5 μM IBA and 0.1% activated charcoal. Over 93% of plantlets survived following acclimatization and transfer to a mixture of sand and vermiculite (1:1, v/v) in trays. PMID:27090564

  11. [Microscopic anatomy of abnormal structure in root tuber of Pueraria lobata].

    PubMed

    Duan, Hai-yan; Cheng, Ming-en; Peng, Hua-sheng; Zhang, He-ting; Zhao, Yu-jiao

    2015-11-01

    Puerariae Lobatae Radix, also known as Gegen, is a root derived from Pueraria lobata. Based on field investigation and the developmental anatomy of root tuber, we have elucidated the relationship between the growth of root tuber and the anomalous structure. The results of analysis showed that the root system of P. lobata was developed from seed and adventitious root and there existed root tuber, adventitious root and conductive root according to morphology and function. The root tuber was developed from adventitious root, its secondary structure conformed to the secondary structure of dicotyledon's root. With the development of root, the secondary phloem of root tuber appeared abnormal vascular tissue, which was distributed like ring in the outside of secondary vascular tissue. The root tuber might have 4-6 concentric circular permutation abnormal vascular tissuelobate, and was formed by the internal development of abnormal vascular tissue. The xylem and phloem of abnormal vascular tissue were the main body of the root tuber. The results reveal the abnormal anatomical structure development of P. lobata, also provides the theoretical basis for reasonable harvest medicinal parts and promoting sustainable utilization of resources of P. lobata. PMID:27097408

  12. Synthetic virus seeds for improved vaccine safety: Genetic reconstruction of poliovirus seeds for a PER.C6 cell based inactivated poliovirus vaccine.

    PubMed

    Sanders, Barbara P; Edo-Matas, Diana; Papic, Natasa; Schuitemaker, Hanneke; Custers, Jerome H H V

    2015-10-13

    Safety of vaccines can be compromised by contamination with adventitious agents. One potential source of adventitious agents is a vaccine seed, typically derived from historic clinical isolates with poorly defined origins. Here we generated synthetic poliovirus seeds derived from chemically synthesized DNA plasmids encoding the sequence of wild-type poliovirus strains used in marketed inactivated poliovirus vaccines. The synthetic strains were phenotypically identical to wild-type polioviruses as shown by equivalent infectious titers in culture supernatant and antigenic content, even when infection cultures are scaled up to 10-25L bioreactors. Moreover, the synthetic seeds were genetically stable upon extended passaging on the PER.C6 cell culture platform. Use of synthetic seeds produced on the serum-free PER.C6 cell platform ensures a perfectly documented seed history and maximum control over starting materials. It provides an opportunity to maximize vaccine safety which increases the prospect of a vaccine end product that is free from adventitious agents.

  13. [Aortic aneurysm in Erdheim's idiopathic cystic medial necrosis in autopsy and forensic medical practice].

    PubMed

    Dolzhansky, O V; Shilova, M A; Pal'tseva, E M; Fedorov, D N; Kocharyan, E Z; Pigolkin, Yu I

    2016-01-01

    дгезии-1 (ICAM-1). Результаты. При морфологическом исследовании наблюдались разрушение ламеллярных единиц, фиброз, медионекроз и фрагментация эластических волокон. Данные признаки оценивали по степени выраженности. При иммуногистохимическом исследовании коллагенов I и III типов определяли пучки хаотично расположенных волокон. Разнонаправленная ориентация гладкомышечных клеток подтверждалась по экспрессии гладкомышечного актина. В гладкомышечных клетках медии и в адвентиции аорты отмечалась выраженная экспрессия маркера апоптоза р53. Экспрессия ФНОα выявлялась в клетках воспалительного инфильтрата стенки аневризмы, ICAM-1 - в эндотелии аорты, vasa vasorum, а также в клетках воспалительного инфильтрата в медии и адвентиции. В работе предложена конструкция патологоанатомического и судебно-медицинского диагнозов, отражающая звенья патогенеза и танатогенеза при идиопатическом кистозном медионекрозе Эрдгейма, а также позволяющая провести шифровку заболевания по МКБ-10. Заключение. Представлены основные патогистологические признаки аневризмы аорты при идиопатическом кистозном ме

  14. Exogenous supplementation of Activin A enhances germ cell differentiation of human embryonic stem cells.

    PubMed

    Duggal, Galbha; Heindryckx, Björn; Warrier, Sharat; Taelman, Jasin; Van der Jeught, Margot; Deforce, Dieter; Chuva de Sousa Lopes, Susana; De Sutter, Petra

    2015-05-01

    Human embryonic stem cells (hESCs) derived in the presence of Activin A (ActA) demonstrate an increased differentiation propensity toward the germ cell lineage. In addition, mouse epiblast stem cells and mouse epiblast-like cells are poised toward germ cell differentiation and are derived in the presence of ActA. We therefore investigated whether supplementation with ActA enhances in vitro hESC differentiation toward germ cell lineage. ActA up-regulated early primordial germ cell (PGC) genes STELLA/DPPA3 (developmental pluripotency associated 3) and tyrosine kinase receptor cKIT in both ActA-derived and standard-derived hESCs indicating its role in priming hESCs toward the PGC lineage. Indeed, ActA plus bone morphogenic protein 4 (BMP4) strongly increased germ cell differentiation potential of hESCs based on the high expression of late PGC markers DAZL (deleted in azoospermia-like) and VASA/DDX4 (DEAD-box polypeptide 4) at mRNA and protein level. Hence, the combination of ActA with BMP4 provides an additional boost for hESCs to develop into postmigratory germ cells. Together with increased VASA expression in the presence of ActA and BMP4, we also observed up-regulation of endoderm-specific genes GATA4 (GATA binding protein 4) and GATA6. Finally, we were able to further mature these in vitro-derived PGC-like cells (PGCLCs) by culturing them in in vitro maturation (IVM) medium, resulting in the formation of germ cell-like clusters and induction of meiotic gene expression. In conclusion, we demonstrate for the first time a synergism between ActA and BMP4 in facilitating germ cell-directed differentiation of hESCs, which is enhanced by extended culture in IVM medium, as shown by cytoplasmic VASA-expressing PGCLCs. We propose a novel relationship between the endoderm and germ cell lineage during hESC differentiation.

  15. Testicular cell-conditioned medium supports embryonic stem cell differentiation toward germ lineage and to spermatocyte- and oocyte-like cells.

    PubMed

    Shah, Syed M; Saini, Neha; Singh, Manoj K; Manik, Radheysham; Singla, Suresh K; Palta, Prabhat; Chauhan, Manmohan S

    2016-08-01

    Testicular cells are believed to secrete various growth factors that activate signaling pathways finally leading to gametogenesis. In vitro gametogenesis is an obscure but paramountly important task primarily because of paucity of the precursor cells and first trimester gonadal tissues. To overcome these limitations for development of in vitro gametes, the present study was designed to induce differentiation of buffalo embryonic stem (ES) cells into germ lineage cells on stimulation by testicular cell-conditioned medium (TCM), on the basis of the assumption that ES cells have the intrinsic property to differentiate into any cell type and TCM would provide the necessary growth factors for differentiation toward germ cell lineage. For this purpose, buffalo ES cells were differentiated as embryoid bodies (EB) in floating cultures and as monolayer adherent cultures in different doses (10%, 20%, and 40%) of TCM for different culture intervals (4, 8, and 14 days), to identify the optimum dose-and-time period. We observed that 40% TCM dose induces highest expression of primordial germ cell-specific (DAZL, VASA, and PLZF), meiotic (SYCP3, MLH1, TNP1/2, and PRM2), spermatocyte-specific (BOULE and TEKT1), and oocyte-specific genes (GDF9 and ZP2/3) for a culture period of 14 days under both floating and adherent differentiation. Immunocytochemical analysis of EBs and adherent cultures revealed presence of primordial germ cell markers (c-KIT, DAZL, and VASA), meiotic markers (SYCP3, MLH1 and PROTAMINE1), spermatocyte markers (ACROSIN and HAPRIN), and oocyte markers (GDF9 and ZP4), indicating progression into post-meiotic gametogenesis. The detection of germ cell-specific proteins in Day 14 EBs like VASA, GDF9, and ZP4 by Western blotting further confirmed germ lineage differentiation. The significantly lower (P < 0.05) concentration of 5-methyl-2-deoxycytidine in optimally differentiated EBs is suggestive of the process of methylation erasure. Oocyte-like structures

  16. Verbal/social autopsy study helps explain the lack of decrease in neonatal mortality in Niger, 2007–2010

    PubMed Central

    Kalter, Henry D; Yaroh, Asma Gali; Maina, Abdou; Koffi, Alain K; Bensaïd, Khaled; Amouzou, Agbessi; Black, Robert E

    2016-01-01

    Background This study was one of a set of verbal/social autopsy (VASA) investigations undertaken by the WHO/UNICEF–supported Child Health Epidemiology Reference Group to estimate the causes and determinants of neonatal and child deaths in high priority countries. The study objective was to help explain the lack of decrease in neonatal mortality in Niger from 2007 to 2010, a period during which child mortality was decreasing. Methods VASA interviews were conducted of a random sample of 453 neonatal deaths identified by the 2010 Niger National Mortality Survey (NNMS). Causes of death were determined by expert algorithm analysis, and the prevalence of household, community and health system determinants were examined along the continuum of maternal and newborn care, the Pathway to Survival for newborn illnesses, and an extended pathway for maternal complications. The social autopsy findings were compared to available data for survivors from the same cohort collected by the NNMS and the 2012 Niger Demographic and Health Survey. Findings Severe neonatal infection and birth asphyxia were the leading causes of early neonatal death in the community and facilities. Death in the community after delayed careseeking for severe infection predominated during the late neonatal period. The levels of nearly all demographic, antenatal and delivery care factors were in the direction of risk for the VASA study decedents. They more often resided rurally (P < 0.001) and their mothers were less educated (P = 0.03) and gave birth when younger (P = 0.03) than survivors’ mothers. Their mothers also were less likely to receive quality antenatal care (P < 0.001), skilled attendance at birth (P = 0.03) or to deliver in an institution (P < 0.001). Nearly half suffered an obstetric complication, with more maternal infection (17.9% vs 0.2%), antepartum hemorrhage (12.5% vs 0.5%) and eclampsia/preeclampsia (9.5% vs 1.6%) than for all births in Niger. Their mothers also

  17. Annual variations of carbonaceous PM2.5 in Malaysia: influence by Indonesian peatland fires

    NASA Astrophysics Data System (ADS)

    Fujii, Y.; Tohno, S.; Amil, N.; Latif, M. T.; Oda, M.; Matsumoto, J.; Mizohata, A.

    2015-08-01

    In this study, we quantified carbonaceous PM2.5 in Malaysia through annual observations of PM2.5, focusing on organic compounds derived from biomass burning. We determined organic carbon (OC), elemental carbon (EC) and concentrations of solvent-extractable organic compounds (biomarkers derived from biomass burning sources and n-alkanes). We observed seasonal variations in the concentrations of pyrolyzed OC (OP), levoglucosan (LG), mannosan (MN), galactosan, syringaldehyde, vanillic acid (VA) and cholesterol. The average concentrations of OP, LG, MN, galactosan, VA and cholesterol were higher during the southwest monsoon season (June-September) than during the northeast monsoon season (December-March), and these differences were statistically significant. Conversely, the syringaldehyde concentration during the southwest monsoon season was lower. The PM2.5 OP/OC4 mass ratio allowed distinguishing the seven samples, which have been affected by the Indonesian peatland fires (IPFs). In addition, we observed significant differences in the concentrations between the IPF and other samples of many chemical species. Thus, the chemical characteristics of PM2.5 in Malaysia appeared to be significantly influenced by IPFs during the southwest monsoon season. Furthermore, we evaluated two indicators, the vanillic acid/syringic acid (VA/SA) and LG/MN mass ratios, which have been suggested as indicators of IPFs. The LG/MN mass ratio ranged from 14 to 22 in the IPF samples and from 11 to 31 in the other samples. Thus, the respective variation ranges partially overlapped. Consequently, this ratio did not satisfactorily reflect the effects of IPFs in Malaysia. In contrast, the VA/SA mass ratio may serve as a good indicator, since it significantly differed between the IPF and other samples. However, the OP/OC4 mass ratio provided more remarkable differences than the VA/SA mass ratio, offering an even better indicator. Finally, we extracted biomass burning emissions' sources such as IPF

  18. Quantitative autoradiography of angiotensin II receptors in brain and kidney: focus on cardiovascular implications

    SciTech Connect

    Gehlert, D.R.; Speth, R.C.; Wamsley, J.K.

    1985-01-01

    Quantitative techniques of receptor autoradiography have been applied to localize (/sup 125/I)-angiotensin II binding sites in brain and kidney. High densities of autoradiographic grains, indicating the presence of angiotensin II receptors, have been localized to several rat brain nuclei including the dorsal motor nucleus of the vagus, nucleus of the solitary tract, anterior pituitary, locus coeruleus and several hypothalamic nuclei. Cat thoracic spinal cord exhibited a high density of sites over the intermedio-lateral cell column. In sections of rat kidney, angiotensin II receptors were detected in the glomerulus, vasa recta and ureter. The cardiovascular implications of these results are apparent and relate angiotensin II to hypertensive mechanisms. Thus, angiotensin II represents an endocoid which is involved in control of blood pressure through its effects on peripheral organs as well as the central nervous system.

  19. Effect of diet on male reproductive tract of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae).

    PubMed

    Lemos, W P; Serráo, J E; Ramalho, F S; Cola Zanuincio, J C; Lacerda, M C

    2005-02-01

    The morphology and histology of the reproductive tract of males of the predator Podisus nigrispinus (Dallas) fed on different diets were studied. P. nigrispinus was fed on diets of: larvae of Alabama argillacea (Hübner), Tenebrio molitor L., Musca domestica L., and an artificial diet. The male reproductive tract, independent of diet, showed testes with intense red coloration in a compact, circular, or slightly oval structure. The vasa deferentia were similar in color to the testes and formed long filaments, which joined with the yellow-cream colored ejaculatory duct. The morphological characteristics of the male reproductive tract were similar under all diets, except for the artificial one. The histological studies demonstrated that independent of the diet the testes of P. nigrispinus were composed of four to six follicles. The testes with six follicles generally had four developed and two atrophied follicles. The morphological and histological differences of the testes of P. nigrispinus when fed with different prey are presented and discussed.

  20. High prevalence of proposed Müllerian duct remnant cysts on the spermatic duct in wild Eurasian otters (Lutra lutra) from Sweden.

    PubMed

    Roos, Anna M; Ågren, Erik O

    2013-01-01

    The spermatic ducts (vasa deferentia) of 235 otters (Lutra lutra) found dead between 1999 and 2012 in Sweden were examined for presence of paraductular cysts. Single or multiple elongated uni- or bilateral cysts parallel to the spermatic duct were noted in 72% of the examined males. The cysts were adjacent to, but did not communicate with the lumen of the spermatic duct, and were usually located within a few centimeters of the testis and epididymis. The cysts are proposed to be congenital Müllerian duct remnants. Other morphologic abnormalities in the reproductive organs were not noted within this study. Possible causes of the incomplete regression of the embryonic female gonadal duct are exposure to environmental contaminants such as elevated concentrations of estrogen-like compounds (endocrine disrupting chemicals), inbreeding, or a naturally occurring anatomic defect. No obvious geographical pattern was observed for otters wi