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Sample records for bovine pericardium tissue

  1. Biomimetic acellular detoxified glutaraldehyde cross-linked bovine pericardium for tissue engineering.

    PubMed

    Mathapati, Santosh; Bishi, Dillip Kumar; Guhathakurta, Soma; Cherian, Kotturathu Mammen; Venugopal, Jayarama Reddy; Ramakrishna, Seeram; Verma, Rama Shanker

    2013-04-01

    Glutaraldehyde (GLUT) processing, cellular antigens, calcium ions in circulation, and phospholipids present in the native tissue are predominantly responsible for calcification, degeneration, and lack of natural microenvironment for host progenitor cell migration in tissue implants. The study presents an improved methodology for adhesion and proliferation of endothelial progenitor cells (EPCs) without significant changes in biomechanical and biodegradation properties of the processed acellular bovine pericardium. The anti-calcification potential of the processed tissue was enhanced by detoxification of GLUT-cross-linked bovine pericardium by decellularization, pretreating it with ethanol or removing the free aldehydes by citric acid treatment and lyophilization. The treated tissues were assessed for biomechanical properties, GLUT ligand quantification, adhesion, proliferation of EPCs, and biodegradability. The results indicate that there was no significant change in biomechanical properties and biodegradability when enzymatic hydrolysis (p>0.05) is employed in detoxified acellular GLUT cross-linked tissue (DBP-G-CA-ET), compared with the native detoxified GLUT cross-linked bovine pericardium (NBP-G-CA-ET). DBP-G-CA-ET exhibited a significant (p>0.05) increase in the viability of EPCs and cell adhesion as compared to acellular GLUT cross-linked bovine pericardium (p<0.05). Lyophilized acellular detoxified GLUT cross-linked bovine pericardium, employed in our study as an alternative to conventional GLUT cross-linked bovine pericardium, might provide longer durability and better biocompatibility, and reduce calcification. The developed bovine pericardium patches could be used in cardiac reconstruction and repair, arteriotomy, soft tissue repair, and general surgical procedures with tissue regeneration dimensions. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Polyethylene glycol-grafted bovine pericardium: a novel hybrid tissue resistant to calcification.

    PubMed

    Vasudev, S C; Chandy, T

    1999-02-01

    Calcification is a frequent cause of the clinical failure of bioprosthetic heart valves fabricated from glutaraldehyde pretreated bovine pericardium (GATBP). An investigation was made of the grafting of different molecular weight polyethylene glycol (PEG 600, 1500, 4000 and 6000) via glutaraldehyde (GA) linkages to bovine pericardium (BP) and of their stability and calcification. The process of the calcification profile was studied by in vitro experiments via incubating pericardial samples in a metastable solution of calcium phosphate. Calcification of bovine pericardium grafted with PEG 6000 was significantly decreased compared to low molecular weight PEG grafts or Sodium dodecyl sulphate- (SDS) and GA-treated tissues. The mechanical properties of these modified tissues after enzyme (Trypsin) digestion and calcification were investigated. The biocompatibility aspects of grafted tissues were also established by monitoring the platelet adhesion, octane contact angle and water of hydration. PEG 6000-grafted tissues retained the maximum strength in trypsin buffer and calcium phosphate solutions. Scanning electron micrographs revealed that the PEG-grafted bovine pericardium had substantially inhibited the platelet-surface attachment and their spreading. It is conceivable that high molecular weight polyethylene glycol-grafted pericardium (a hybrid tissue) may be a suitable calcium-resistant material for developing prosthetic valves due to their stability and biocompatibility.

  3. Immunoproteomic identification of bovine pericardium xenoantigens

    PubMed Central

    Griffiths, Leigh G.; Choe, Leila H.; Reardon, Kenneth F.; Dow, Steven W.; Orton, E. Christopher

    2008-01-01

    Bovine pericardium is an important biomaterial with current application in glutaraldehyde-fixed bioprosthetic heart valves and possible future application as an unfixed biological scaffold for tissue engineering. The importance of both humoral and cell-mediated rejection responses toward fixed and unfixed xenogeneic tissues has become increasingly apparent. However, the full scope and specific identities of bovine pericardium proteins that can elicit an immune response remain largely unknown. In this study, an immunoproteomic approach was used to survey bovine pericardium proteins for their ability to elicit a humoral immune response in rabbits. A two-stage protein extraction protocol was used to separate bovine pericardium proteins into water- and lipid-soluble fractions. Two-dimensional gel electrophoresis was performed to separate the proteins from each fraction. Western blots were generated from two-dimensional gels of both bovine pericardium protein fractions. These blots were probed with serum from rabbits immunized with bovine pericardium and a secondary antibody was used to assess for IgG positivity. Western blots were compared to duplicate two-dimensional gels and proteins in matched spots were identified by tandem mass spectrometry. Thirty-one putative protein antigens were identified, eight of which are known to be antigenic from previous studies. All of the putative antigens demonstrated progressive staining intensity with increasing days of post-exposure serum. Identified antigenic proteins represented a variety of functional and structural protein types, and included both cellular and matrix proteins. The results of this study have implications for the use of bovine pericardium as a biomaterial in bioprostheses and tissue engineering applications, as well as xenotransplantation in general. PMID:18514307

  4. Protein extraction and 2-DE of water- and lipid-soluble proteins from bovine pericardium, a low-cellularity tissue.

    PubMed

    Griffiths, Leigh G; Choe, Leila; Lee, Kelvin H; Reardon, Kenneth F; Orton, E Christopher

    2008-11-01

    Bovine pericardium (BP) is an important biomaterial used in the production of glutaraldehyde-fixed heart valves and tissue-engineering applications. The ability to perform proteomic analysis on BP is useful for a range of studies, including investigation of immune rejection after implantation. However, proteomic analysis of fibrous tissues such as BP is challenging due to their relative low-cellularity and abundance of extracellular matrix. A variety of methods for tissue treatment, protein extraction, and fractionation were investigated with the aim of producing high-quality 2-DE gels for both water- and lipid-soluble BP proteins. Extraction of water-soluble proteins with 3-(benzyldimethylammonio)-propanesulfonate followed by n-dodecyl beta-D-maltoside extraction and ethanol precipitation for lipid-soluble proteins provided the best combination of yield, spot number, and resolution on 2-DE gels (Protocol E2). ESI-quadrupole/ion trap or MALDI-TOF/TOF MS protein identifications were performed to confirm bovine origin and appropriate subcellular prefractionation of resolved proteins. Twenty-five unique, predominantly cytoplasmic bovine proteins were identified from the water-soluble fraction. Thirty-two unique, predominantly membrane bovine proteins were identified from the lipid-soluble fraction. These results demonstrated that the final protocol produced high-quality proteomic data from this important tissue for both cytoplasmic and membrane proteins.

  5. Preseeding of human vascular cells in decellularized bovine pericardium scaffold for tissue-engineered heart valve: an in vitro and in vivo feasibility study.

    PubMed

    Yang, Min; Chen, Chang-Zhi; Shu, Yu-Sheng; Shi, Wei-Ping; Cheng, Shao-Fei; Gu, Y John

    2012-08-01

    Human vascular cells from saphenous veins have been used for cell seeding on the synthetic scaffolds for constructing tissue-engineered heart valve (TEHV). However, little is known about the seeding of human vascular cells on bovine pericardium, a potential natural scaffold for TEHV. This study was aimed to assess the basic in vitro and in vivo characteristics of the human vascular cells seeded on decellularized bovine pericardium. In vitro, bovine pericardium samples with cell seeding were inspected on day 7, 14, and 21 by histology, scanning electron microscopy, and immunohistochemistry. In vivo, experiments were performed in nude mice by bilateral dorsal incision for the implantation of decellularized bovine pericardium with and without cell seeding. Results demonstrated that a total of 8-10 × 10(6) cells were obtained within 4-5 wk by the primary co-culture, which were detected positive for von Willebrand factor, α-smooth muscle actin antibodies, and fibronectin, indicating the presence of endothelial cells, smooth muscle cells, and fibroblasts, respectively. In vitro, the seeded cells showed a steady increase of endothelial activity from day 1 to day 7 and remained stable until day 21. After 30 days of implantation in vivo, the cells on the decellularized bovine pericardium could differentiate directionally and show all the identities of human endothelial cells, smooth muscle cells, and fibroblasts. These results indicate that the human vascular cells from the saphenous vein are an optional cell source for seeding on decellularized bovine pericardium scaffold for constructing TEHV. Copyright © 2012 Wiley Periodicals, Inc.

  6. Glutaraldehyde Treatment Elicits Toxic Response Compared to Decellularization in Bovine Pericardium

    PubMed Central

    Umashankar, P.R.; Mohanan, P.V.; Kumari, T.V.

    2012-01-01

    Glutaraldehyde-stabilized bovine pericardium is used for clinical application since 1970s because of its desirable features such as less immunogenicity and acceptable durability. However, a propensity for calcification is reported on account of glutaraldehyde treatment. In this study, commercially available glutaraldehyde cross-linked bovine pericardium was evaluated for its in vitro cytotoxic effect, macrophage activation, and in vivo toxic response in comparison to decellularized bovine pericardium. Glutaraldehyde-treated bovine pericardium and its extract were observed to be cytotoxic and it also caused significant inflammatory cytokine release from activated macrophages. Significant antibody response, calcification response, necrotic, and inflammatory response were noticed in glutaraldehyde-treated bovine pericardium in comparison to decellularized bovine pericardium in a rat subcutaneous implantation model. Glutaraldehyde-treated bovine pericardium also failed in acute systemic toxicity testing and intracutaneous irritation testing as per ISO 10993. With respect to healing and implant remodeling, total lack of host tissue incorporation and angiogenesis was noticed in glutaraldehyde-treated bovine pericardium compared to excellent host fibroblast incorporation and angiogenesis within the implant in decellularized bovine pericardium. In conclusion, using in vitro and in vivo techniques, this study has demonstrated that glutaraldehyde-treated bovine pericardium elicits toxic response compared to decellularized bovine pericardium which is not congenial for long-term implant performance. PMID:22736904

  7. Epidural Bovine Pericardium Facilitates Dissection During Cranioplasty: A Technical Note.

    PubMed

    Griessenauer, Christoph J; He, Lucy; Salem, Mohamed; Chua, Michelle; Ogilvy, Christopher S; Thomas, Ajith J

    2015-12-01

    Adhesions and scarring of the subcutaneous tissue to the dura mater or dural substitute often complicate cranioplasty. We present our experience with epidural bovine pericardium as a barrier membrane to minimize adhesions and facilitate separation of tissue layers. A cohort of patients who underwent decompressive craniectomy and subsequent cranioplasty at a major academic institution in the United States from August 2007 to October 2013 and had epidural bovine pericardium placed as a barrier membrane was retrospectively reviewed. Medical records and imaging studies were reviewed for a number of variables including presence of adhesions, infection, contusions, and operative complications. Twenty-nine patients (male-to-female = 1:1.1; mean age 45 ± 14.7 years) who underwent decompressive craniectomy with placement of epidural bovine pericardium with subsequent cranioplasty were identified. The median interval between craniectomy and cranioplasty was 64 days, and autologous bone was used for cranioplasty in 86.2% of cases. The average size of cranial defect was 71.2 ± 28.5 cm(2). At the time of cranioplasty, no or minimal adhesions were found between the subcutaneous tissue and the epidural bovine pericardium. There were 2 (6.9%) infections, 2 (6.9%) patients had contusion after the cranioplasty, and no patient had a complication after cranioplasty that required reoperation. Epidural bovine pericardium at the time of decompressive craniectomy facilitates dissection at the time of cranioplasty and is not associated with any additional risks. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Care during freeze-drying of bovine pericardium tissue to be used as a biomaterial: a comparative study.

    PubMed

    Polak, Roberta; Pitombo, Ronaldo N M

    2011-10-01

    Bovine pericardium (BP) tissue is widely used in the manufacture of bioprosthetics. The effects of freeze-drying on the BP tissue have been studied by some researchers in order to decrease their cytotoxicity due to preservation in formaldehyde solution, and to increase the lifetime of the product in storage. This study was undertaken in order to study the effect of freeze-drying in the structure of BP. To perform this study BP samples were freeze-dried in two different types of freeze-dryers available in our laboratory: a laboratory freeze-dryer, in which it was not possible to control parameters and a pilot freeze-dryer, wherein all parameters during freezing and drying were controlled. After freeze-drying processes, samples were analyzed by SEM, Raman spectroscopy, tensile strength, water uptake tests and TEM. In summary, it has been demonstrated that damages occur in collagen fibers by the loss of bulk water of collagen structure implicating in a drastic decreasing of BP mechanical properties due to its structural alterations. Moreover, it was proven that the collagen fibrils suffered breakage at some points, which can be attributed to the uncontrolled parameters during drying.

  9. Nanofibers coated on acellular tissue-engineered bovine pericardium supports differentiation of mesenchymal stem cells into endothelial cells for tissue engineering.

    PubMed

    Mathapati, Santosh; Bishi, Dillip Kumar; Venugopal, Jayarama Reddy; Cherian, Kotturathu Mammen; Guhathakurta, Soma; Ramakrishna, Seeram; Verma, Rama Shanker

    2014-04-01

    This study aimed to develop biodegradable, polymer-based nanofibers coated on acellular tissue-engineered bovine pericardium (ATEBP) for cell interfaces, enabling more exquisite functionality, such as mesenchymal stem cell (MSC) adhesion, proliferation and differentiation into endothelial cells for tissue engineering. ATEBP coated with nanofibers of poly(L-lactic acid)-co-poly(ε-caprolactone) (PLACL) and a blend of PLACL and gelatin were analyzed for human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells. The cell culture-based approach showed an increase in human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells on ATEBP coated with PLACL/gelatin nanofibers compared with ATEBP and PLACL nanofibers coated on ATEBP. ATEBP coated with PLACL/gelatin nanofibrous scaffolds, along with human bone marrow-derived MSCs differentiated into endothelial cells, might improve the scaffolds' functionality for tissue engineering.

  10. Short duration gluteraldehyde cross linking of decellularized bovine pericardium improves biological response.

    PubMed

    Umashankar, P R; Arun, T; Kumari, T V

    2011-06-01

    Gluteraldehyde stabilized bovine pericardium is used for clinical application since 1970s because of its desirable features such as less immunogenicity and acceptable durability. However, a propensity for calcification and long term implant failure is reported because of gluteraldehyde treatment. There is also failure of implant to integrate into host tissue because of its resistance to tissue remodeling. Decellularized bovine pericardium, a potential alternative allows tissue remodeling but it has problems such as immunogenicity and chronic inflammatory response. In this study, decellularized bovine pericardium was subjected to short duration, low concentration gluteraldehyde cross-linking at two levels and its biological response (both in vitro and in vivo) was compared with un-crosslinked decellularized bovine pericardium and fully crosslinked normal bovine pericardium. It was observed that both un-crosslinked and partially crosslinked decellularized bovine pericardium to be non-cytotoxic and it caused significantly less inflammatory cytokine release such as TNF alpha and IL1beta from activated macrophages. Among all groups, short duration 0.2% Gluteraldehyde treated decellularized bovine pericardium showed significantly less antibody response and inflammatory response compared to un-crosslinked decellularized pericardium, short duration 0.6% gluteraldehyde treated decellularized bovine pericardium or completely cross linked bovine pericardium in juvenile rat subcutaneous implantation model. Moreover, short duration 0.2% gluteraldehyde crosslinked decellularized bovine pericardium showed minimum calcification, better host fibroblast incorporation, new collagen deposition and angiogenesis within the implant. These attributes may finally lead to better implant remodeling and sustained implant function during clinical use. Copyright © 2011 Wiley Periodicals, Inc.

  11. Age dependent differences in collagen alignment of glutaraldehyde fixed bovine pericardium.

    PubMed

    Sizeland, Katie H; Wells, Hannah C; Higgins, John; Cunanan, Crystal M; Kirby, Nigel; Hawley, Adrian; Mudie, Stephen T; Haverkamp, Richard G

    2014-01-01

    Bovine pericardium is used for heart valve leaflet replacement where the strength and thinness are critical properties. Pericardium from neonatal animals (4-7 days old) is advantageously thinner and is considered as an alternative to that from adult animals. Here, the structures of adult and neonatal bovine pericardium tissues fixed with glutaraldehyde are characterized by synchrotron-based small angle X-ray scattering (SAXS) and compared with the mechanical properties of these materials. Significant differences are observed between adult and neonatal tissue. The glutaraldehyde fixed neonatal tissue has a higher modulus of elasticity (83.7 MPa) than adult pericardium (33.5 MPa) and a higher normalised ultimate tensile strength (32.9 MPa) than adult pericardium (19.1 MPa). Measured edge on to the tissue, the collagen in neonatal pericardium is significantly more aligned (orientation index (OI) 0.78) than that in adult pericardium (OI 0.62). There is no difference in the fibril diameter between neonatal and adult pericardium. It is shown that high alignment in the plane of the tissue provides the mechanism for the increased strength of the neonatal material. The superior strength of neonatal compared with adult tissue supports the use of neonatal bovine pericardium in heterografts.

  12. Age Dependent Differences in Collagen Alignment of Glutaraldehyde Fixed Bovine Pericardium

    PubMed Central

    Sizeland, Katie H.; Wells, Hannah C.; Higgins, John; Cunanan, Crystal M.; Kirby, Nigel; Hawley, Adrian; Mudie, Stephen T.; Haverkamp, Richard G.

    2014-01-01

    Bovine pericardium is used for heart valve leaflet replacement where the strength and thinness are critical properties. Pericardium from neonatal animals (4–7 days old) is advantageously thinner and is considered as an alternative to that from adult animals. Here, the structures of adult and neonatal bovine pericardium tissues fixed with glutaraldehyde are characterized by synchrotron-based small angle X-ray scattering (SAXS) and compared with the mechanical properties of these materials. Significant differences are observed between adult and neonatal tissue. The glutaraldehyde fixed neonatal tissue has a higher modulus of elasticity (83.7 MPa) than adult pericardium (33.5 MPa) and a higher normalised ultimate tensile strength (32.9 MPa) than adult pericardium (19.1 MPa). Measured edge on to the tissue, the collagen in neonatal pericardium is significantly more aligned (orientation index (OI) 0.78) than that in adult pericardium (OI 0.62). There is no difference in the fibril diameter between neonatal and adult pericardium. It is shown that high alignment in the plane of the tissue provides the mechanism for the increased strength of the neonatal material. The superior strength of neonatal compared with adult tissue supports the use of neonatal bovine pericardium in heterografts. PMID:25295250

  13. An evaluation of Admedus' tissue engineering process-treated (ADAPT) bovine pericardium patch (CardioCel) for the repair of cardiac and vascular defects.

    PubMed

    Strange, Geoff; Brizard, Christian; Karl, Tom R; Neethling, Leon

    2015-03-01

    Tissue engineers have been seeking the 'Holy Grail' solution to calcification and cytotoxicity of implanted tissue for decades. Tissues with all of the desired qualities for surgical repair of congenital heart disease (CHD) are lacking. An anti-calcification tissue engineering process (ADAPT TEP) has been developed and applied to bovine pericardium (BP) tissue (CardioCel, AdmedusRegen Pty Ltd, Perth, WA, Australia) to eliminate cytotoxicity, improve resistance to acute and chronic inflammation, reduce calcification and facilitate controlled tissue remodeling. Clinical data in pediatric patients, and additional pre-market authorized prescriber data demonstrate that CardioCel performs extremely well in the short term and is safe and effective for a range of congenital heart deformations. These data are supported by animal studies which have shown no more than normal physiologic levels of calcification, with good durability, biocompatibility and controlled healing.

  14. Improved biocompatibility by postfixation treatment of aldehyde fixed bovine pericardium.

    PubMed

    Moritz, A; Grimm, M; Eybl, E; Grabenwöger, M; Windberger, U; Dock, W; Böck, P; Wolner, E

    1990-01-01

    Long-standing release of locally cytotoxic aldehyde concentrations is responsible for lack of spontaneous endothelialization and increased calcification of glutaraldehyde fixed bovine pericardium. Postfixation treatment with amino acids made in vitro endothelialization of bioprosthetic heart valves possible. Such treated pericardium calcified significantly less (13 +/- 4 micrograms/mg dry weight) than did conventionally processed pericardium (114 +/- 25 micrograms/mg) after 63 days of subcutaneous implantation in rats. To test the ability for spontaneous in vivo endothelialization, 5 sheep had 6 mm grafts made from postfixation treated pericardium (PTP) implanted into the carotid artery, compared to PTFE grafts on the contralateral side, which spontaneously endothelialize in animal models. In a pregnant animal, both grafts occluded. All remaining pericardial grafts remained patent, but one additional PTFE graft occluded and another one was stenosed. The area covered with red thrombus was significantly smaller in the PTP grafts (3.05 +/- 3.9%) than in the PTFE grafts 42 +/- 14% (p = 0.0036); TEM and SEM showed endothelial cells growing directly on the PTP, but only on myofibroblasts in PTFE grafts. Postfixation treatment of glutaraldehyde fixed pericardium aids spontaneous endothelialization and decreases tissue calcification.

  15. Corporoplasty using bovine pericardium grafts in complex penile prosthesis implantation surgery.

    PubMed

    Lopes, Eduardo J A; Kuwano, Andre Y; Guimaraes, Andreia N; Flores, Jesuino P; Jacobino, Modesto A O

    2009-01-01

    This paper is the first, to our knowledge, to propose the use of a bovine pericardium graft in corporoplasty for penile prosthesis implantation. The advantages of bovine pericardium graft have been demonstrated by its use in cardiac surgery, including low cost, biocompatibility, impermeability, resistance to dilatation, flexibility, low likelihood of retraction, absence of antigenic reaction and natural absorption of the tissue. In this paper, we propose the use of this heterologous material graft in corporoplasty for penile prosthesis implantation. Five patients with a history of erosion, infection and fibrosis, mean time of follow-up 32 months (range 9-48 months). Bovine pericardium was used to cover large areas of implanted penile prostheses when use of the tunica albuginea was unfeasible. The surgical procedure resulted in no complications in all patients. Bovine pericardium may substitute synthetic and autologous material with the additional advantages of lower cost and greater availability.

  16. PORCINE VENA CAVA AS AN ALTERNATIVE TO BOVINE PERICARDIUM IN BIOPROSTHETIC PERCUTANEOUS HEART VALVES

    PubMed Central

    Munnelly, Amy; Cochrane, Leonard; Leong, Joshua; Vyavahare, Naren

    2011-01-01

    Percutaneous heart valves are revolutionizing valve replacement surgery by offering a less invasive treatment option for high-risk patient populations who have previously been denied the traditional open chest procedure. Percutaneous valves need to be crimped to accommodate a small-diameter catheter during deployment, and they must then open to the size of heart valve. Thus the material used must be strong and possess elastic recoil for this application. Most percutaneous valves utilize bovine pericardium as a material of choice. One possible method to reduce the device delivery diameter is to utilize a thin, highly elastic tissue. Here we investigated porcine vena cava as an alternative to bovine pericardium for percutaneous valve application. We compared the structural, mechanical, and in vivo properties of porcine vena cava to those of bovine pericardium. While the extracellular matrix fibers of pericardium are randomly oriented, the vena cava contains highly aligned collagen and elastin fibers that impart strength to the vessel in the circumferential direction and elasticity in the longitudinal direction. Moreover, the vena cava contains a greater proportion of elastin, whereas the pericardium matrix is mainly composed of collagen. Due to its high elastin content, the vena cava is significantly less stiff than the pericardium, even after crosslinking with glutaraldehyde. Furthermore, the vena cava’s mechanical compliance is preserved after compression under forces similar to those exerted by a stent, whereas pericardium is significantly stiffened by this process. Bovine pericardium also showed surface cracks observed by scanning electron microscopy after crimping that were not seen in vena cava tissue. Additionally, the vena cava exhibited reduced calcification (46.64 ± 8.15 μg Ca/mg tissue) as compared to the pericardium (86.79 ± 10.34 μg/mg). These results suggest that the vena cava may enhance leaflet flexibility, tissue resilience, and tissue

  17. Effect of chronic inflammation and immune response on regeneration induced by decellularized bovine pericardium.

    PubMed

    Umashankar, P R; Arun, T; Kumary, T V

    2013-08-01

    Decellularised tissue produces a variety of host responses ranging from constructive remodeling to scarring on account of its differences in the source of tissue, processing or sterilization methods. In this study, in vivo regeneration induced by decellularised bovine pericardium with or without mild glutaraldehyde crosslinking was studied in relation to its immune and inflammatory response using rat abdominal regeneration model. Mild glutaraldehyde crosslinking was done to subdue inflammatory and immune response without compromising host cell incorporation and graft remodeling. Evaluations were done at both 21 and 90 days. Un-crosslinked decellularised bovine pericardium showed more intense macrophage response predominantly of M2 phenotype at 90 days indicating chronic inflammatory response compared to mildly crosslinked group. This group also showed significant increase in plasma cell and lymphocyte count indicating immune stimulation. Lymphocyte transformation test detected presence of bovine pericardial antigen sensitized lymphocytes at both periods in un-crosslinked group. Lymphocytes from mildly crosslinked group failed to respond in this test at both periods. Significantly higher antibody response was also noted at both periods in un-crosslinked group. However, abdominal wall regeneration was observed only in animals implanted with un-crosslinked decellularised bovine pericardium at 90 days. From the above findings, it is inferred that un-crosslinked decellularised bovine pericardium produced significant chronic inflammatory response at 90 days and stimulated both humoral and cell mediated immune response in comparison to mildly crosslinked decellularised bovine pericardium. Yet this group produced skeletal muscle formation within graft at 90 days. Copyright © 2012 Wiley Periodicals, Inc.

  18. Extraction and characterization of highly purified collagen from bovine pericardium for potential bioengineering applications.

    PubMed

    Santos, Maria Helena; Silva, Rafael M; Dumont, Vitor C; Neves, Juliana S; Mansur, Herman S; Heneine, Luiz Guilherme D

    2013-03-01

    Bovine pericardium is widely used as a raw material in bioengineering as a source of collagen, a fundamental structural molecule. The physical, chemical, and biocompatibility characteristics of these natural fibers enable their broad use in several areas of the health sciences. For these applications, it is important to obtain collagen of the highest possible purity. The lack of a method to produce these pure biocompatible materials using simple and economically feasible techniques presents a major challenge to their production on an industrial scale. This study aimed to extract, purify, and characterize the type I collagen protein originating from bovine pericardium, considered to be an abundant tissue resource. The pericardium tissue was collected from male animals at slaughter age. Pieces of bovine pericardium were enzymatically digested, followed by a novel protocol developed for protein purification using ion-exchange chromatography. The material was extensively characterized by electrophoresis, scanning electron microscopy, energy dispersive X-ray spectroscopy, and infrared spectroscopy. The results showed a purified material with morphological properties and chemical functionalities compatible with type I collagen and similar to a highly purified commercial collagen. Thus, an innovative and relatively simple processing method was developed to extract and purify type I collagen from bovine tissue with potential applications as a biomaterial for regenerative tissue engineering. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Porcine vena cava as an alternative to bovine pericardium in bioprosthetic percutaneous heart valves.

    PubMed

    Munnelly, Amy E; Cochrane, Leonard; Leong, Joshua; Vyavahare, Naren R

    2012-01-01

    Percutaneous heart valves are revolutionizing valve replacement surgery by offering a less invasive treatment option for high-risk patient populations who have previously been denied the traditional open chest procedure. Percutaneous valves need to be crimped to accommodate a small-diameter catheter during deployment, and they must then open to the size of heart valve. Thus the material used must be strong and possess elastic recoil for this application. Most percutaneous valves utilize bovine pericardium as a material of choice. One possible method to reduce the device delivery diameter is to utilize a thin, highly elastic tissue. Here we investigated porcine vena cava as an alternative to bovine pericardium for percutaneous valve application. We compared the structural, mechanical, and in vivo properties of porcine vena cava to those of bovine pericardium. While the extracellular matrix fibers of pericardium are randomly oriented, the vena cava contains highly aligned collagen and elastin fibers that impart strength to the vessel in the circumferential direction and elasticity in the longitudinal direction. Moreover, the vena cava contains a greater proportion of elastin, whereas the pericardium matrix is mainly composed of collagen. Due to its high elastin content, the vena cava is significantly less stiff than the pericardium, even after crosslinking with glutaraldehyde. Furthermore, the vena cava's mechanical compliance is preserved after compression under forces similar to those exerted by a stent, whereas pericardium is significantly stiffened by this process. Bovine pericardium also showed surface cracks observed by scanning electron microscopy after crimping that were not seen in vena cava tissue. Additionally, the vena cava exhibited reduced calcification (46.64 ± 8.15 μg Ca/mg tissue) as compared to the pericardium (86.79 ± 10.34 μg/mg). These results suggest that the vena cava may provide enhanced leaflet flexibility, tissue resilience, and tissue

  20. Cuspal dehiscence at a post and along the stent cloth in a bovine pericardium heart valve implanted for seven years.

    PubMed

    Guidoin, Robert; Bes, Taniela Marli; Cianciulli, Tomas Francisco; Klein, Julianne; Li, Bin; Gauvin, Robert; Guzman, Randolph; Rochette-Drouin, Olivier; Germain, Lucie; Zhang, Ze

    2012-01-01

    A failing mitral valve prosthesis made from bovine pericardium was explanted from a 50-year-old patient. Preoperative transthoracic-echocardiography had confirmed severe mitral regurgitation due to structural failure of this HP Bio bovine pericardium heart valve prosthesis. The explanted device was examined macroscopically, by scanning electron microscopy (SEM), by light microscopy, and by transmission electron microscopy (TEM). Samples of unassembled patches of bovine pericardium were used as a pre-implantation control to better understand the changes that occurred in the structure of the pericardium following the 7 years of implantation. Examination confirmed complete dehiscence of a cusp along a valve post and the stent: This detached cusp was observed floating in the bloodstream at echocardiography. The fibrous pannus overgrowth was well developed along the stent and extended to the bottom of the cusps both on the inflow and the outflow sides. The fibrous panni were found to be poorly adhesive to the pericardium cusps and had become stiff, thus impairing the opening and closure of the valve. The structure of the pericardium cusps was severely deteriorated compared to the control bovine pericardium tissue samples. The collagen bundles were frequently broken and more stretched in the explanted device, lacking the wavy histological pattern of normal collagen fibers. However, the tissues were devoid of any calcification. In conclusion, the failure mode of this valve was the dehiscence of a cusp from a valve post and along the stent cloth in the absence calcification.

  1. A comparative study of bovine and porcine pericardium to highlight their potential advantages to manufacture percutaneous cardiovascular implants.

    PubMed

    Gauvin, Robert; Marinov, Georgi; Mehri, Yayhe; Klein, Julianne; Li, Bin; Larouche, Danielle; Guzman, Randolph; Zhang, Ze; Germain, Lucie; Guidoin, Robert

    2013-11-01

    Prosthetic heart valves designed to be implanted percutaneously must be loaded within delivery catheters whose diameter can be as low as 18 F (6 mm). This mandatory crimping of the devices may result in deleterious damages to the tissues used for valve manufacturing. As bovine and porcine pericardial tissue are currently given preference because of their excellent availability and traceability, a preliminary comparative study was undertaken to highlight their potential advantages. Bovine and pericardium patches were compared morphologically (light microscopy, scanning electron microscopy and transmission electron microscopy). The acute thrombogenicity of both materials was measured in term of platelet uptake and observed by scanning electron microscopy, porcine intact and injured arteries being used as controls. The pericardium specimens were also subjected to uniaxial tensile tests to compare their respective mechanical characteristics. Both pericardiums showed a layered architecture of collagen bundles presenting some interstitial cells. They displayed wavy crimps typical of an unloaded collagenous tissue. The collagen bundles were not bound together and the fibrils were parallel with characteristic periodicity patterns of cross striations. The mesothelial cells found in vivo on the serous surface were no longer present due to tissue processing, but the adjacent structure was far more compacted when compared to the fibrous side. The fibrinocollagenous surfaces were found to be more thrombogenic for both bovine and porcine tissues and the serous side of the porcine pericardium retained more platelets when compared to the bovine samples, making the acute thrombogenicity more important in the porcine pericardium. Both bovine and porcine pericardium used in cardiovascular implantology can be selected to manufacture percutaneous heart valves. The selection of one pericardium preferably to the other should deserve additional testing regarding the innocuousness of

  2. [STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS].

    PubMed

    Xiao, Hongtao; Tian, Shemin; Zha, Xinjian; Wei, Ying; Huang, Hongjun; Li, Yun; Yang, Huanna; Xia, Chengde; Niu, Xihua

    2015-10-01

    To investigate the effects of modification of acellular bovine pericardium with 1-ethyl-3-(3- dinethylami-nopropyl) carbodimide (EDC)/N-hydroxysuccininide (NHS) or genipin and find out the best crosslinking reagent. The cellular components of the bovine pericardiums were removed. The effects of decellularization were tested by HE staining. The acellular bovine pericardiums were crosslinked with EDC/NHS (EDC/NHS group) or genipin (genipin group). The properties of the crosslinked acellular matrix were evaluated by scanning electron microscope (SEM), matrix thickness, crosslinking index, mechanical property, denaturation temperature, enzymatic degradation, and cytotoxicity test before and after the crosslinking. Acellular bovine pericardium (ABP group) or normal bovine pericardium (control group) were harvested as controls. SEM showed that collagen fibers were reticulated in bovine pericardial tissues after crosslinked by EDC/NHS or genipin, and relative aperture of the collagen fiber was from 10 to 20 μm. The thickness and denaturation temperature of the scaffolds were increased significantly after crosslinking with EDC/NHS or genipin (P < 0.05), while there was no significant difference between EDC/NHS group and genipin group (P > 0.05). The difference had no statistical significance in crosslinking index between EDC/NHS group and genipin group (t = 0.205, P = 0.218). The degradation rate in EDC/NHS group and genipin group was significantly lower than that in ABP group and control group (P < 0.05). Elastic modulus and fracture stress in EDC/NHS group and genipin group were significantly lower than those in ABP group (P < 0.05), but there was no significant difference among EDC/NHS group, genipin group, and control group (P > 0.05). The break elongation in EDC/NHS group and genipin group were significantly increased than those in ABP group and control group (P < 0.05). The difference had no statistical significance in stability and mechanical properties between EDC

  3. Mechanical loading of bovine pericardium accelerates enzymatic degradation.

    PubMed

    Ellsmere, J C; Khanna, R A; Lee, J M

    1999-06-01

    Bioprosthetic heart valves fail as the result of two simultaneous processes: structural deterioration and calcification. Leaflet deterioration and perforation have been correlated with regions of highest stress in the tissue. The failures have long been assumed to be due to simple mechanical fatigue of the collagen fibre architecture; however, we have hypothesized that local stresses-and particularly dynamic stresses-accelerate local proteolysis, leading to tissue failure. This study addresses that hypothesis. Using a novel, custom-built microtensile culture system, strips of bovine pericardium were subjected to static and dynamic loads while being exposed to solutions of microbial collagenase or trypsin (a non-specific proteolytic enzyme). The time to extend to 30% strain (defined here as time to failure) was recorded. After failure, the percentage of collagen solubilized was calculated based on the amount of hydroxyproline present in solution. All data were analyzed by analysis of variance (ANOVA). In collagenase, exposure to static load significantly decreased the time to failure (P < 0.002) due to increased mean rate of collagen solubilization. Importantly, specimens exposed to collagenase and dynamic load failed faster than those exposed to collagenase under the same average static load (P = 0.02). In trypsin, by contrast, static load never led to failure and produced only minimal degradation. Under dynamic load, however, specimens exposed to collagenase, trypsin, and even Tris/CaCl2 buffer solution, all failed. Only samples exposed to Hanks' physiological solution did not fail. Failure of the specimens exposed to trypsin and Tris/CaCl2 suggests that the non-collagenous components and the calcium-dependent proteolytic enzymes present in pericardial tissue may play roles in the pathogenesis of bioprosthetic heart valve degeneration.

  4. The effect of different treatment modalities on the calcification potential and cross-linking stability of bovine pericardium.

    PubMed

    van den Heever, J J; Neethling, W M L; Smit, F E; Litthauer, D; Joubert, G

    2013-03-01

    Porcine heart valves and bovine pericardium exhibit suitable properties for use as substitutes in cardiothoracic surgery, but must meet several requirements to be safe and efficient. Treatment with glutaraldehyde (GA) render some of these requirements, but calcification and degradation post-implant remain a problem. This study aimed to identify additional biochemical treatments that will minimize calcification potential without compromising the physical properties of pericardium. Pericardium treated with GA calcified severely after 8 weeks in the subcutaneous rat model, compared to tissue treated with higher concentrations of glycosaminoglycans (GAG) and commercial Glycar patches. GA, lower concentrations GAG and Glycar pericardium had high denaturation temperatures due to enhanced cross-linking. Tensile strength of GA tissue was significantly lower than GAG-treated or Glycar tissues, due to lower water content with resultant lower flexibility and suppleness. Pericardium treated with 0.01 M GAG gave acceptable denaturation temperatures, tensile strength and reduced calcification potential. All tissue treatments evoked comparable host immune responses, and no significant difference in resistance to enzymatic degradation. Ineffective stabilization and fixation of cross-links following GAG treatment, as well as limited penetration into the pericardium, resulted in GAG leaching out into the surrounding host tissue or storage medium, and prohibits safe clinical use of such tissue.

  5. The antithrombotic versus calcium antagonistic effects of polyethylene glycol grafted bovine pericardium.

    PubMed

    Vasudev, S C; Chandy, T; Sharma, C P

    1999-07-01

    Cardiovascular calcification, the formation of calcium phosphate deposits in cardiovascular tissue, is a common end stage phenomenon affecting a wide variety of bioprosthesis. This study proposes a novel approach of reducing pericardial calcification and thrombosis via coupling polyethylene glycols (PEG) to glutaraldehyde treated bovine pericardium via acetal linkages. The calcification of the PEG modified tissue and the control pericardium (extracted and glutaraldehyde treated) was investigated by in vivo rat subcutaneous implantation models and by in vitro meta stable calcium phosphate solutions. Scanning electron microscopy showed that calcification primarily involved the surface of collagen fibrils and the intrafibrillar spaces. However, the grafting of pericardium with PEG-20,000 had dramatically modified the surface and subsequently inhibited the deposits of calcium. Further, the modified tissue had also reduced the platelet surface attachment. Such a reduced calcification of PEG modified tissues can be explained by decrease of free aldehyde groups, a space filling effect and therefore improved biostability and synergistic blood compatible effects of PEG after coupling to the tissues. This simple method can be a useful anticalcification treatment for implantable tissue valves.

  6. Mapping the calcification of bovine pericardium in rat model by enhanced micro-computed tomography.

    PubMed

    Liu, Jing; Zhong, Shengping; Lan, Hualin; Meng, Xu; Zhang, Haibo; Fan, Yubo; Wang, Yuxing; Wang, Chunren; Wang, Zhaoxu

    2014-09-01

    The calcification initiation and progression of bioprosthetic heart valve were investigated in a rat model by enhanced micro-computed tomography, together with histologic study and scanning electron microscope analysis. The implantation data at early stage showed apparent dendritic patterns in the radiographic images for the glutaraldehyde-treated bovine pericardium and this dendritic pattern was verified to be associated with the vessel distribution in the tissue. Histologic study and scanning electron microscope analysis both indicated that the calcium deposits in the pericardium vessels regions were more grievous than those scattered in the collagen fibers in the first two weeks after implantation. Subsequently, calcification spreaded and the entire sample was severely calcified in 60 days.

  7. Photo-oxidized bovine pericardium in congenital cardiac surgery: single-centre experience.

    PubMed

    Baird, Christopher W; Myers, Patrick O; Piekarski, Breanna; Borisuk, Michele; Majeed, Amara; Emani, Sitaram M; Sanders, Stephen P; Nathan, Meena; Del Nido, Pedro J

    2017-02-01

    Dye-mediated photo-oxidation of pericardium is an alternative method to chemical treatment with glutaraldehyde for cross-linking collagen, providing biostability of the patch material while avoiding late calcification and cytotoxicity. There are few data available, on using photo-oxidation-treated pericardium, in congenital cardiac surgery. This study reports the outcomes using Photofix™ bovine pericardium in neonates, infants, children and young adults undergoing paediatric cardiac surgery. A total of 490 patches in 383 consecutive operations (364 patients) were used in the surgical repair of congenital heart defects at our institution from October 2008 to October 2011. Recorded variables included demographic data, age at operation, primary cardiac diagnosis, associated complications and number, type and location of patches placed and patch-related reintervention. Median age at operation was 5.3 years, ranging from <1 month to 56 years. The overall survival rate at late follow-up was 92%, and no deaths were related to failure of the tissue substitute. Two patients (0.5%) underwent reintervention late due to patch material failure: one for residual shunt after Rastelli repair and one for aneurysmal dilatation of a right ventricular outflow tract patch. The patch material was explanted in 8 patients at a mean of 20 months (range, 1-72 months) following implantation. Histological examination revealed mild to moderate inflammation with variable calcification. Photo-oxidized bovine pericardium demonstrated excellent performance when used as a patch material in cardiovascular repair in children. Its handling characteristics and biocompatibility are consistent with a wide range of applications.

  8. Preparation and characterization of an acellular bovine pericardium intended for manufacture of valve bioprostheses.

    PubMed

    Goissis, Gilberto; Giglioti, Aparecida de Fátima; Braile, Domingo Marcolino

    2011-05-01

    Major problems with biological heart valves post-implantation are associated with progressive structural deterioration and calcification attributed to glutaraldehyde processing, dead cells, and cell fragments present in the native tissue. In spite of these problems, glutaraldehyde still is the reagent of choice. The results with acellular matrix xenograft usually prepared by detergent treatment in association with enzymes are rather conflicting because while preserving mechanical properties, tissue morphology and collagen structure are process dependent. This work describes a chemical approach for the preparation of an acellular bovine pericardium matrix intended for the manufacture of heart valve bioprostheses. Cell removal was performed by an alkaline extraction in the presence of calcium salts for periods ranging from 6 to 48 h. The results showed that cell removal was achieved after 12 h, with swelling and negative charge increasing with processing time. Nevertheless, collagen fibril structure, ability to form fibrils, and stability to collagenase were progressive after 24-h processing. There was no denaturation of the collagen matrix. A process is described for the preparation of acellular bovine pericardium matrices with preserved fibril structure and morphology for the manufacture of cardiac valve bioprostheses and may be used in other applications for tissue reconstruction.

  9. Combined anticalcification treatment of bovine pericardium with decellularization and hyaluronic acid derivative.

    PubMed

    Zhu, Deyi; Jin, Liqiang; Wang, Xuemei; Xu, Li; Liu, Tianqi

    2014-01-01

    The objective of this work was to evaluate the effect of decellularization and hyaluronic acid derivative on the improvement of anticalcification of glutaraldehyde fixed bovine pericardium (GFBP) using a rat subcutaneous implantation model A cell extraction process was employed to remove the cells and cellular components from bovine pericardium (BP), leaving a framework of largely insoluble collagen. Then acellular BP was cross-linked by glutaraldehyde solution and treated with hyaluronic acid derivative (HA-ADH) which was obtained by coupling adipic dihydrazide (ADH) on-COOH of hyaluronic acid (HA). The results of in vivo calcification tests showed that the calcium content was decreased dramatically by decellularization alone (from 28.07 ± 18.87 to 2.44 ± 0.55 μg Ca/mg dry tissue after 8 weeks' implantation), and even less concentration was shown by the combination of HA derivative treatment and decellularization (GFaBP-HA group) (0.25 ± 0.08 μg Ca/mg dry tissue after 8 weeks' implantation). In addition, GFaBP-HA group not only presented a lower degree of calcification, but also showed lower ratios of Ca/P molar, which corresponded to amorphous calcium phosphates. The obtained results indicated that GFaBP-HA was a potential candidate for the manufacture of anticalcification bioprostheses.

  10. Use of bovine pericardium graft for abdominal wall reconstruction in contaminated fields

    PubMed Central

    D’Ambra, Luigi; Berti, Stefano; Feleppa, Cosimo; Magistrelli, Prospero; Bonfante, Pierfrancesco; Falco, Emilio

    2012-01-01

    AIM: To employ, in such conditions, a biological graft such as bovine pericardium that offers resistance to infection. METHODS: In our surgical department, from January 2006 to June 2010, 48 patients underwent abdominal wall reconstruction using acellular bovine pericardium; of these 34 patients had a contaminated wound due to diffuse peritonitis (complicated diverticulitis, bowel perforation, intestinal infarction, strangled hernia, etc.) and 14 patients had hernia relapse on infected synthetic mesh. RESULTS: In our series, one patient died of multi-organ failure 3 d after surgery. After placement of the pericardium mesh four cases of hernia relapse occurred. CONCLUSION: Recurrence rate is similar to that of prosthetic mesh repair and the application of acellular bovine pericardium (Tutomesh®, Tutogen Medical Gmbh Germany) is moreover a safe and feasible option that can be employed to manage complicated abdominal wall defects where prosthetic mesh is unsuitable. PMID:22905285

  11. Alkaline phosphatase activity of glutaraldehyde-treated bovine pericardium used in bioprosthetic cardiac valves.

    PubMed

    Maranto, A R; Schoen, F J

    1988-10-01

    Bioprosthetic valves fail frequently because of pathological mineralization, a process that begins in cell remnants of the glutaraldehyde (GLUT) fixed tissue. Other pathological cardiovascular calcification and physiological mineralization in skeletal/dental tissues are both largely initiated in cell-derived membranous structures (often called "matrix vesicles"), and the enzyme alkaline phosphatase (AP) likely has an important function in the pathogenesis of mineral nucleation. This study tested the hypothesis that AP might also be present in and contribute to calcification of bioprosthetic valves. AP activity of fresh and GLUT-treated bovine pericardium was measured by the conversion of p-nitrophenyl phosphate to p-nitrophenol. Following 24 hours in 0.6% HEPES-buffered GLUT and storage for 2 weeks in 0.2% GLUT, considerable AP hydrolytic activity remained in GLUT-treated tissue relative to that of fresh tissue (Vmax, 24 vs. 45 mumol reaction product/min/mg tissue protein, respectively), although binding was somewhat reduced (Km, 1.9 X 10(3) vs. 1.4 X 10(3) microM substrate, respectively). Enzyme reaction product was demonstrated in both fixed and fresh tissue by light microscopic histochemical studies, confirming the biochemical results. Reaction product was noted along membranes of vascular endothelial cells and interstitial fibroblasts, the sites of early calcific deposits in bioprosthetic valves, by ultrastructural examination of GLUT-treated tissue. We conclude that GLUT-treated bovine pericardium retains much of the hydrolytic activity of AP, an enzyme associated with normal skeletal and pathological cardiovascular and noncardiovascular mineralization, and suggest that further examination of the mechanistic role of this enzyme may stimulate new approaches for slowing or preventing calcification of bioprosthetic tissue.

  12. Conjunctival structural and functional reconstruction using acellular bovine pericardium graft (Normal GEN®) in rabbits.

    PubMed

    Huang, Danping; Xu, Bing; Yang, Xiaonan; Xu, Binbin; Zhao, Jing

    2016-04-01

    To evaluate the effectiveness of acellular bovine pericardium grafts (Normal GEN®) used as scaffolds for conjunctival reconstruction. The acellular bovine pericardium graft and the amnion graft were implanted into the bulbar conjunctival defects of adult rabbits. Conjunctival samples of implanted materials and blank defect controls were observed at day 3, 7, 14, 21, 28, and 56 postoperatively. Histological examination was observed at day 14, 28, and 56 of surgery, including hematoxylin-eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining, while immunofluorescent microscopy was observed at 14 days and 28 days after surgery. Results were compared among the Normal GEN®, amnion, and blank defect controls. All three groups showed complete conjunctival reconstruction. Wounds that were not grafted closed by formation of conjunctival scar characterized by a linear array of densely packed collagen fibers in Tenon's capsule. Subepithelial tissue in the grafted groups comprised a loosely organized network of randomly oriented collagen that resembled that of the normal bulbar conjunctiva. However, there was a dense layer of aligned collagen between the conjunctival Tenon's capsule and the sclera in the NormalGEN® group, about 250 μm in thickness. Implantation of the NormalGEN® graft promoted the formation of conjunctiva as a kind of scaffold both in structure and in function. It had more advantageous mechanical properties than the amnion, strong and elastic, during the period of conjunctival reconstruction.

  13. Combined application of acellular bovine pericardium and hyaluronic acid in prevention of postoperative pericardial adhesion.

    PubMed

    Shen, Jia; Xu, Zhi Wei

    2014-03-01

    An experiment was designed to find the suitable acellular bovine pericardium (ABP) patch in pericardial cavity reconstruction and to evaluate the effect of sodium hyaluronic acid (NaHA) on inflammatory reaction in prevention of pericardial adhesions. The pericardial adhesion model was established in 20 rabbits, weighing from 3.2 to 3.6 kg. Groups were classified as follows: Group A (n = 5), the control group, the pericardium was directly closed; Group B (n = 5), 0.15% glutaraldehyde-treated ABP (low cross-link degree); Group C, 0.3% glutaraldehyde-treated ABP (middle cross-link degree); Group D, 0.15% glutaraldehyde-treated ABP + NaHA solution. Blood samples were collected at 6 h, 24 h, 3 days, and 5 days, to assay postoperative inflammatory reaction. The tenacity and severity of adhesions were evaluated 2 months after operation, by macroscopic and microscopic examinations, and Q-PCR (real-time quantitative polymerase chain reaction) test was used to quantitatively analyze the associated genes with adhesion. Pericardium regeneration was demonstrated by immunohistochemical technique to identify mesothelial cells. In Group D, the serum concentration of tumor necrosis factor-α (TNF-α) was significantly lower in the early postoperative period, and the mean adhesion score (adhesion between the epicardium and ABP) was significantly lower compared with the control group (Groups D vs. A: 0.20 ± 0.45 vs. 2.00 ± 0.71, P = 0.009*). The signs of degradation of the ABPs were observed 2 months postoperation in Groups D and B. Immunohistochemically, the positive cytokeratin AE1 staining results demonstrated the relatively total regeneration of the pericardium in Group D. Signs of regeneration were observed in Group D. Compared with the control group, the level of TGF-β2 in Group D was significantly lower (0.00132 ± 0.00114, P = 0.022*). The TGF-β3 level was statistically significant, being highest in Group D (0.00805 ± 0.00136, P = 0.029*). The mean quantity of Smad6 in

  14. Mechanical assessment of bovine pericardium using Müeller matrix imaging, enhanced backscattering and digital image correlation analysis

    PubMed Central

    Cuando-Espitia, Natanael; Sánchez-Arévalo, Francisco; Hernández-Cordero, Juan

    2015-01-01

    Mechanical characterization of tissue is an important but complex task. We demonstrate the simultaneous use of Mueller matrix imaging (MMI), enhanced backscattering (EBS) and digital image correlation (DIC) in a bovine pericardium (BP) tensile test. The interest in BP relies on its wide use as valve replacement and biological patch. We show that the mean free path (MFP), obtained through EBS measurements, can be used as an indicator of the anisotropy of the fiber ensemble. Our results further show a good correlation between retardance images and displacement vector fields, which are intrinsically related with the fiber interaction within the tissue. PMID:26309759

  15. Treatment of an Iatrogenic Left Internal Mammary Artery to Pulmonary Artery Fistula with a Bovine Pericardium Covered Stent

    SciTech Connect

    Heper, Gulumser Barcin, Cem; Iyisoy, Atila; Tore, Hasan F.

    2006-10-15

    We report a case with an acquired fistula between the left internal mammary artery and the pulmonary artery following coronary bypass surgery treated with a bovine pericardium covered stent. We also reviewed similar cases reported previously.

  16. Transplantation material bovine pericardium: biomechanical and immunogenic characteristics after decellularization vs. glutaraldehyde-fixing.

    PubMed

    Hülsmann, Jörn; Grün, Katja; El Amouri, Sonya; Barth, Mareike; Hornung, Katrin; Holzfuß, Carlheinz; Lichtenberg, Artur; Akhyari, Payam

    2012-01-01

    Today, bovine pericardium (BP) is extensively investigated as a biomaterial for the generation of various bioimplants. But despite the commercial distribution, and the development of methods either to remove (decellularization) or to mask (chemical cross-linking, for example by glutaraldehyde [GA] treatment) the xenogeneic antigen epitopes, yet questions around the immunogenic reactivity of BP remain. The aim of this study is the comparison of crucial tissue characteristics, that is, biomechanical properties, the presence of αGal epitopes, and residual DNA in acellular vs. GA-fixed BP. Bovine pericardium was either cross-linked with 0.6% GA or decellularized according to two common protocols using either sodium dodecyl sulfate (SDS) and desoxycholic acid (DCA) or trypsin and ethylenediaminetetraacetic acid (EDTA). The resulting extracellular matrix was prone to one-dimensional tensile testing. The tissue content for αGal was evaluated by immunoblotting, and residual DNA was determined by a commercial assay. Untreated BP served as control. In contrast to previous reports, we found a pronounced decrease in the elastic modulus (E-Modulus) for common GA treatment and overall smaller values for the elastic moduli after decellularization (P < 0.05). In parallel, we observed an overall increased ultimate elongation of acellular and cross-linked BP, although ultimate stress values did not significantly differ. SDS/DCA decellularized BP revealed a dramatic reduction in the DNA content and an almost complete removal of αGal epitopes, whereas the trypsin/EDTA protocol retained a residual DNA content of almost 50% and with a great trail of αGal signal. GA-treated tissue had a remarkable content of DNA and αGal. Although chemically fixated BP is clinically still in wide use, for example, for biological heart valve engineering, our results suggest that an improved biomaterial preparation may be provided by appropriate decellularization. SDS/DCA decellularized BP shows

  17. Effects of glutaraldehyde concentration and fixation time on material characteristics and calcification of bovine pericardium: implications for the optimal method of fixation of autologous pericardium used for cardiovascular surgery.

    PubMed

    Lee, Cheul; Lim, Hong-Gook; Lee, Chang-Ha; Kim, Yong Jin

    2017-03-01

    Autologous pericardium, which is widely used in the field of cardiovascular surgery, is usually fixed with glutaraldehyde (GA) to improve handling and provide biomechanical stability. However, an optimal method of GA fixation of autologous pericardium is not known. The objective of this study was to evaluate the effects of GA concentration and fixation time on material characteristics and calcification of bovine pericardium. Bovine pericardial tissues were fixed with different concentrations of GA (0.3, 0.4, 0.5 and 0.6%) for different exposure times (10 and 20 min). Material characteristics of the fixed tissues were assessed by mechanical test, thermal stability test and pronase test. The tissues were subcutaneously implanted into 3-week-old rats for 2 months, and the calcium contents of the explanted tissues were measured. Differences between the groups were evaluated by two-way analysis of variance. Differently treated tissues showed no significant differences in tensile strength. The mean elongation at break of the pericardial tissues fixed with 0.5 and 0.6% was significantly higher compared with 0.3 and 0.4% when fixed for 20 min. The mean elongation at break of the pericardial tissues fixed for 20 min was significantly higher compared with 10 min when fixed with 0.5 and 0.6%. Thermal stability test revealed significantly higher mean shrinkage temperature of the pericardial tissues fixed with 0.6% compared with lower concentrations irrespective of fixation time. The mean shrinkage temperature of the pericardial tissues fixed for 20 min was significantly higher compared with 10 min irrespective of GA concentration. Pronase test revealed significantly lower mean percent remaining weight of the pericardial tissues fixed with 0.3% compared with higher concentrations irrespective of fixation time. The mean percent remaining weight of the pericardial tissues fixed for 20 min was significantly higher compared with 10 min irrespective of GA

  18. Cytotoxicity of PVPAC-treated bovine pericardium: a potential replacement for glutaraldehyde in biological heart valves.

    PubMed

    Barros, Janaina Aline Galvão; Filippin-Monteiro, Fabiola Branco; de Oliveira, Edson Mendes; Campa, Ana; Catalani, Luiz Henrique; Pitombo, Ronaldo de Nogueira Moraes; Polakiewicz, Bronislaw

    2014-04-01

    Acellular biological tissues, including bovine pericardium (BP), have been proposed as biomaterial for tissue engineering. BP is usually modified chemically to improve mechanical and biological properties using glutaraldehyde, the standard reagent for preservation of fresh bioprosthetic materials. Glutaraldehyde-fixed BP (Glut-BP), the most widely used material in heart valve manufacture, has been associated with calcification in vivo. In an attempt to reduce this issue and maintain its biocompatibility, this study assesses the physical properties and cytotoxicity of lyophilized BP treated with poly (vinylpyrrolidone-co-acrolein) (PVPAC-BP), a novel copolymer, as a substitute for glutaraldehyde. For that, PVPAC-BP surface ultrastructure, elastic function, water uptake and tissue calcification were evaluated. For the analysis of biocompatibility, fibroblasts (3T3-L1) and endothelial cells (HUVEC) were cultured on PVPAC-BP, Untreated-BP and Glut-BP. Nitric oxide (NO) release assay, fluorescence and SEM images of endothelial cells adhered on scaffolds were also performed. As results, the data show some advantages of PVPAC-BP over the Glut-BP. The PVPAC-BP maintains partially the original ultrastructure and elastic properties, improves scaffold hydration, and presents less calcium phosphate deposits. The cells demonstrated strong attachment, high proliferation rate, and formation of a monolayer on PVPAC-BP. Attached cells were also able to release NO de-monstrating regular metabolism. In conclusion, PVPAC may be considered as a promising alternative to BP treatment improving the efficiency of cell attachment and proliferation and also avoid immunogenicity.

  19. Ten-year comparative analysis of bovine pericardium and autogenous vein for patch angioplasty in patients undergoing carotid endarterectomy.

    PubMed

    Kim, Ji-Hoon; Cho, Yong-Pil; Kwon, Tae-Won; Kim, Hyangkyoung; Kim, Geun-Eun

    2012-04-01

    To evaluate early and late clinical outcomes of carotid endarterectomy (CEA) with bovine pericardium patch in comparison with autogenous vein. During a 10-year period, 456 CEAs were performed using patch closure of the arteriotomy with bovine pericardium (252 cases) and autogenous vein (204 cases). Retrospectively, surgical outcomes were evaluated and compared regarding CEA-related parameters, early and late mortality and morbidity rates, and the incidence of restenosis and aneurysmal dilatation between patients with bovine pericardium patch closure and those with autogenous vein closure. The two groups were comparable regarding basic demographics, clinical data, and anatomic data, except the incidence of coronary or peripheral arterial diseases. In patients with bovine pericardium patch closure, the total operating time and carotid clamping duration were statistically significantly shorter than in those with autogenous vein closure (P < 0.01). During the early postoperative period, 10 major (stroke and death) complications (2.2%) occurred without statistically significant difference between the two groups. The incidence of early minor postoperative complications was less with bovine pericardium patch closure (5.6% vs. 10.8%; P < 0.05). With a mean follow-up of 62 months for bovine pericardium patch closure and 67 months for autogenous vein closure, the incidence of restenosis was similar, but aneurysmal dilatation was higher in patients with autogenous vein closure with a statistically significant difference (0% vs. 2.0%; P < 0.05). CEA with bovine pericardium patch angioplasty showed excellent early and late clinical outcomes. Our results demonstrated bovine pericardium to be a suitable patch material for routine use in CEA. Copyright © 2012 Annals of Vascular Surgery Inc. Published by Elsevier Inc. All rights reserved.

  20. Decellularization and Delipidation Protocols of Bovine Bone and Pericardium for Bone Grafting and Guided Bone Regeneration Procedures

    PubMed Central

    Ferroni, Letizia; Guazzo, Riccardo; Sbricoli, Luca; De Benedictis, Giulia; Finotti, Luca; Isola, Maurizio; Bressan, Eriberto; Zavan, Barbara

    2015-01-01

    The combination of bone grafting materials with guided bone regeneration (GBR) membranes seems to provide promising results to restore bone defects in dental clinical practice. In the first part of this work, a novel protocol for decellularization and delipidation of bovine bone, based on multiple steps of thermal shock, washes with detergent and dehydration with alcohol, is described. This protocol is more effective in removal of cellular materials, and shows superior biocompatibility compared to other three methods tested in this study. Furthermore, histological and morphological analyses confirm the maintenance of an intact bone extracellular matrix (ECM). In vitro and in vivo experiments evidence osteoinductive and osteoconductive properties of the produced scaffold, respectively. In the second part of this study, two methods of bovine pericardium decellularization are compared. The osmotic shock-based protocol gives better results in terms of removal of cell components, biocompatibility, maintenance of native ECM structure, and host tissue reaction, in respect to the freeze/thaw method. Overall, the results of this study demonstrate the characterization of a novel protocol for the decellularization of bovine bone to be used as bone graft, and the acquisition of a method to produce a pericardium membrane suitable for GBR applications. PMID:26191793

  1. Decellularization and Delipidation Protocols of Bovine Bone and Pericardium for Bone Grafting and Guided Bone Regeneration Procedures.

    PubMed

    Gardin, Chiara; Ricci, Sara; Ferroni, Letizia; Guazzo, Riccardo; Sbricoli, Luca; De Benedictis, Giulia; Finotti, Luca; Isola, Maurizio; Bressan, Eriberto; Zavan, Barbara

    2015-01-01

    The combination of bone grafting materials with guided bone regeneration (GBR) membranes seems to provide promising results to restore bone defects in dental clinical practice. In the first part of this work, a novel protocol for decellularization and delipidation of bovine bone, based on multiple steps of thermal shock, washes with detergent and dehydration with alcohol, is described. This protocol is more effective in removal of cellular materials, and shows superior biocompatibility compared to other three methods tested in this study. Furthermore, histological and morphological analyses confirm the maintenance of an intact bone extracellular matrix (ECM). In vitro and in vivo experiments evidence osteoinductive and osteoconductive properties of the produced scaffold, respectively. In the second part of this study, two methods of bovine pericardium decellularization are compared. The osmotic shock-based protocol gives better results in terms of removal of cell components, biocompatibility, maintenance of native ECM structure, and host tissue reaction, in respect to the freeze/thaw method. Overall, the results of this study demonstrate the characterization of a novel protocol for the decellularization of bovine bone to be used as bone graft, and the acquisition of a method to produce a pericardium membrane suitable for GBR applications.

  2. Results of Carotid Endarterectomy Using Bovine Pericardium Patch Closure, with a Review of Pertinent Literature.

    PubMed

    Olsen, Stephen B; Mcquinn, William C; Feliciano, Philip

    2016-03-01

    Patch closure after carotid endarterectomy lowers risk of recurrent stenosis and perioperative neurologic events. Although vein patches have theoretic benefits, few differences between vein and nonautologous patches have been noted. Bovine pericardium has been increasingly used in carotid surgery over the last 15 years, with favorable reports. This study reviews a large carotid surgery series using this patch. Data were gathered on 464 consecutive carotid operations using the bovine pericardium patch; all complications were analyzed and pertinent literature review was performed. Of 464 procedures, 218 (47%) were performed for symptomatic disease. Total perioperative ipsilateral stroke rate was 0.65 per cent. A low rate of postoperative bleeding was noted. Recurrence of >70 per cent stenosis was 1.6 per cent at follow-up of 26 months in 410 patients. Handling was subjectively described as excellent. Literature review comprising 3239 carotid operations revealed low risks of stroke, bleeding requiring intervention, or high rates of recurrent stenosis. Bovine pericardium patch angioplasty has low perioperative and late complications and good handling characteristics, making it an acceptable choice for use in carotid surgery.

  3. Tear and decohesion of bovine pericardial tissue.

    PubMed

    Tobaruela, Almudena; Elices, Manuel; Bourges, Jean Yves; Rojo, Francisco Javier; Atienza, José Miguel; Guinea, Gustavo

    2016-10-01

    The aim of this study was to evaluate quantitatively the fracture-by tear and delamination-of bovine pericardium tissues which are usually employed for the manufacture of bioprosthetic valves. A large number of samples (77) were tested in root-to-apex and circumferential directions, according to a standardised tear test (ASTM D 1938). Before performing the tear test, some samples were subjected to 1000 cycles of fatigue to a maximum stress of 3MPa. Fracture toughness of tearing and delamination were computed by following a simple fracture model. The study showed significantly lower values of delamination toughness compared with tear delamination. Moreover, tear forces were different in each test direction, revealing a clear orthotropic behaviour. All these results, as well as the testing procedure, could be of value for future research in the physiological function of pericardium tissues and clinical applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Bovine pericardium in treating large corneal perforation secondary to alkali injury: A case report

    PubMed Central

    Mokhtar, Ezanee

    2008-01-01

    To describe use of a locally processed bovine pericardium (BP) to cover a large central corneal perforation following alkali injury and discuss postoperative outcome. A 27-year-old Malay male patient presented two weeks after alkali splashed in his left eye while working. A clinical diagnosis of left central corneal ulcer with limbal ischemia following alkali injury with secondary infection was made. After failed medical therapy, we performed a Gunderson conjunctival flap under local anesthesia that retracted after one week and resulted in a large central corneal perforation with surrounding stromal thinning. The perforation was covered with a locally processed BP xenograft (Lyolemb) supplied by the National Tissue Bank, University Sains Malaysia. Nine months follow-up showed a well-taken graft without any exposure/dehiscence and minimal inflammation. Amniotic membrane transplantation when used as a patch graft needs an urgent tectonic graft to promote corneal stability in patients with severe corneal thinning. The use of processed BP can be a viable option in treating such cases. PMID:18711278

  5. Decellularization as an anticalcification method in stentless bovine pericardium valve prosthesis: a study in sheep.

    PubMed

    Collatusso, Claudinei; Roderjan, João Gabriel; Vieira, Eduardo Discher; Myague, Nelson Itiro; Noronha, Lúcia de; Costa, Francisco Diniz Affonso da

    2011-01-01

    The objective was to analyze the decellularization process with SDS in glutaraldehyde-preserved bovine pericardium as an anticalcification method in a circulatory sheep model. The valved tubs were implanted in pulmonary artery position in sheep by 180 days. The animals were divided in two groups of 8 animals: control group--glutaraldehyde-preserved bovine pericardium and the study group--decellularized bovine pericardium with 0,1% SDS and glutaraldehyde-preserved. After explantation the tubs were analized by x-ray macroscopy, hematoxilin-eosin, alizarin-red and Russel-Movatz pentacromic histology. The calcium content was measured by flame atomic absorption spectrometry. There was no early mortality, but two animals in each group died during the study. All cusps in the control group were severely calcified and in some points in the conduits, while the decellularized group did not show macroscopic calcification. Data were proved by x-ray and histologycal exams. The matrix was preserved in histologycal analysis in decellularized group, without gross calcification. The wall conduits calcium content was 35,25 ± 42,13 µg/mg in the control group versus 15,75 ± 10,44 µg/mg in the decellularized one: in the cusps was 264,4 ± 126,16 µg/mg in control group versus 94,29 ± 27,05 µg/mg in decellularized group (P = 0,009). The decellularization with 0.1% SDS was effective as an anticalcification method in bovine pericardial grafts implanted in a sheep circulatory model for 180 days.

  6. Bovine Pericardium Patch Wrapping Intestinal Anastomosis Improves Healing Process and Prevents Leakage in a Pig Model

    PubMed Central

    Testini, Mario; Gurrado, Angela; Portincasa, Piero; Scacco, Salvatore; Marzullo, Andrea; Piccinni, Giuseppe; Lissidini, Germana; Greco, Luigi; De Salvia, Maria Antonietta; Bonfrate, Leonilde; Debellis, Lucantonio; Sardaro, Nicola; Staffieri, Francesco; Carratù, Maria Rosaria; Crovace, Antonio

    2014-01-01

    Failure of intestinal anastomosis is a major complication following abdominal surgery. Biological materials have been introduced as reinforcement of abdominal wall hernia in contaminated setting. An innovative application of biological patch is its use as reinforcement of gastrointestinal anastomosis. The aim of study was to verify whether the bovine pericardium patch improves the healing of anastomosis, when in vivo wrapping the suture line of pig intestinal anastomosis, avoiding leakage in the event of deliberately incomplete suture. Forty-three pigs were randomly divided: Group 1 (control, n = 14): hand-sewn ileo-ileal and colo-colic anastomosis; Group 2 (n = 14): standard anastomosis wrapped by pericardium bovine patch; Group 3 (n = 1) and 4 (n = 14): one suture was deliberately incomplete and also wrapped by patch in the last one. Intraoperative evaluation, histological, biochemical, tensiometric and electrophysiological studies of intestinal specimens were performed at 48 h, 7 and 90 days after. In groups 2 and 4, no leak, stenosis, abscess, peritonitis, mesh displacement or shrinkage were found and adhesion rate decreased compared to control. Biochemical studies showed mitochondrial function improvement in colic wrapped anastomosis. Tensiometric evaluations suggested that the patch preserves the colic contractility similar to the controls. Electrophysiological results demonstrated that the patch also improves the mucosal function restoring almost normal transport properties. Use of pericardium bovine patch as reinforcement of intestinal anastomosis is safe and effective, significantly improving the healing process. Data of prevention of acute peritonitis and leakage in cases of iatrogenic perforation of anastomoses, covered with patch, is unpublished. PMID:24489752

  7. Right ventricular function after repair of tetralogy of Fallot: a comparison between bovine pericardium and porcine small intestinal extracellular matrix.

    PubMed

    Naik, Ronak; Johnson, Jason; Kumar, T K S; Philip, Ranjit; Boston, Umar; Knott-Craig, Christopher J

    2017-05-29

    The porcine small intestinal extracellular matrix reportedly has the potential to differentiate into viable myocardial cells. When used in tetralogy of Fallot repair, it may improve right ventricular function. We evaluated right ventricular function after repair of tetralogy of Fallot with extracellular matrix versus bovine pericardium. Subjects with non-transannular repair of tetralogy of Fallot with at least 1 year of follow-up were selected. The extracellular matrix and bovine pericardium groups were compared. We used three-dimensional right ventricular ejection fraction, right ventricle global longitudinal strain, and tricuspid annular plane systolic excursion to assess right ventricular function. The extracellular matrix group had 11 patients, whereas the bovine pericardium group had 10 patients. No differences between the groups were found regarding sex ratio, age at surgery, and cardiopulmonary bypass time. The follow-up period was 28±12.6 months in the extracellular matrix group and 50.05±17.6 months in the bovine pericardium group (p=0.001). The mean three-dimensional right ventricular ejection fraction (55.7±5.0% versus 55.3±5.2%, p=0.73), right ventricular global longitudinal strain (-18.5±3.0% versus -18.0±2.2%, p=0.44), and tricuspid annular plane systolic excursions (1.59±0.16 versus 1.59±0.2, p=0.93) were similar in the extracellular matrix group and in the bovine pericardium group, respectively. Right ventricular global longitudinal strain in healthy children is reported at -29±3% in literature. In a small cohort of the patients undergoing non-transannular repair of tetralogy of Fallot, there was no significant difference in right ventricular function between groups having extracellular matrix versus bovine pericardium patches followed-up for more than 1 year. Lower right ventricular longitudinal strain noted in both the groups compared to healthy children.

  8. Biocompatibility and calcification of bovine pericardium employed for the construction of cardiac bioprostheses treated with different chemical crosslink methods.

    PubMed

    Jorge-Herrero, Eduardo; Fonseca, Carlos; Barge, Alexandra P; Turnay, Javier; Olmo, Nieves; Fernández, Pilar; Lizarbe, María A; García Páez, José M

    2010-05-01

    The use of biological materials in the construction of bioprostheses requires the application of different chemical procedures to improve the durability of the material without producing any undesirable effects. A number of crosslinking methods have been tested in biological tissues composed mainly of collagen. The aim of this study was to evaluate the in vitro biocompatibility, the mechanical properties, and in vivo calcification of chemically modified bovine pericardium using glutaraldehyde acetals (GAAs) in comparison with glutaraldehyde (GA) treatment. Homsy's tests showed that the most cytotoxic treatment is GA whereas GAA treatments showed lower cytotoxicity. Regarding the mechanical properties of the modified materials, no significant differences in stress at rupture were detected among the different treatments. Zeta-Potential showed higher negative values for GA treatment (-4.9 +/- 0.6 mV) compared with GAA-0.625% (-2.2 +/- 0.5 mV) and GAA-1% (-2.2 +/- 0.4 mV), which presented values similar to native tissue. Similar results were obtained for calcium permeability coefficients which showed the highest values for GA treatment (0.12 +/- 0.02 mm(2)/min), being significantly lower for GAA treatments or non-crosslinked pericardium. These results confirmed the higher propensity of the GA-treated tissues for attraction of calcium cations and were in good agreement with the calcification degree obtained after 60 days implantation into young rats, which was significantly higher for the GA group (22.70 +/- 20.80 mg/g dry tissue) compared with GAA-0.625% and GAA-1% groups (0.49 +/- 0.28 mg/g dry tissue and 3.51 +/- 3.27 mg/g dry tissue, respectively; P < 0.001). In conclusion, GAA treatments can be considered a promising alternative to GA treatment.

  9. Bovine pericardium retail preserved in glutaraldehyde and used as a vascular patch

    PubMed Central

    2011-01-01

    Background In this study we evaluated the performance of bovine pericardium preserved in glutaraldehyde used as a vascular patch. Methods Fourteen young pigs, six females and eight males, weighting 10.3 - 18.4 kg were used in our study. We implanted three remnants in each pig, two in the abdominal aorta and one was juxtaposed to the peritoneum. The smooth face (SF) and rough face (RF) of each remnant were implanted turned to the vessel inner portion and one remnant was juxtaposed to the peritoneum. The animals were sacrificed in 4.5 - 8 months after surgery (75 - 109 kg). The remnants were assessed for aorta wall, fibroses formation in inner apposition and calcification related to the face turned to the vessel inner portion. Results The rough face showed a lower dilatation level compared to the face implanted in adjacent aorta. There was no difference between intensity and/or incidence of graft calcification when the superficies were compared. The bovine pericardium preserved in glutaraldehyde did not show alterations in its structure when implanted with different faces turned to the inner portion of vessel. Conclusion When turned to the inner portion of the vessel, the rough face of the remnant presented a lower dilatation in relation to the adjacent aorta and a better quality of endothelium layer and did not show a difference between intensity and/or incidence of graft calcification. PMID:22192162

  10. Use of bovine pericardium as a wrapping material for hydroxyapatite orbital implants

    PubMed Central

    Gupta, M; Puri, P; Rennie, I G

    2002-01-01

    Aim: To present the results of 27 patients who had enucleation for malignant melanoma of the choroid with hydroxyapatite implant wrapped in bovine pericardium. Method: A retrospective study was performed on 27 patients, 12 males and 15 females, who had enucleation as a primary treatment for their choroidal melanomas. The patients were followed up at 1 week, 1 month, 3 months, and then every 6 months. A conformer was fitted at 1 week and an artificial eye at 1 month. The average follow up was 1.7 years. Results: No patient had extrusion of the implant. One patient needed repair of the wound, two patients required a lateral tarsal strip, and one patient developed a conjunctival granuloma, which did not need excision. In one patient there was shallowing of the inferior fornix. The cosmetic results and ocular movement were satisfactory in all but one patient. Conclusion: Use of bovine pericardium as wrapping material for the hydroxyapatite implants has shown promising results with minimal extrusion rates providing an effective alternative for sclera, eliminating the potential risks of CJD. PMID:11864885

  11. Inhibition of bioprosthesis calcification due to synergistic effect of Fe/Mg ions to polyethylene glycol grafted bovine pericardium.

    PubMed

    Vasudev, S C; Chandy, T; Umasankar, M M; Sharma, C P

    2001-10-01

    Calcification has limited the durability of bioprosthetic heart valves fabricated from glutaraldehyde pretreated porcine aortic valves or bovine pericardium (BP). The present study describes calcium antagonistic effect of polyethylene glycol grafted bovine pericardium (PEG-GABP) with Fe2+/Mg2+ delivery from a co-matrix system in rat subcutaneous model. Retrieved samples were biochemically evaluated for calcification and alkaline phosphate (AP) activity. Scanning electron micrographs of 21-day explants had shown excessive calcification with glutaraldehyde treated BP (control). However, the PEG grafting and Fe/Mg release had substantially inhibited the deposition of calcium on BP. The extractable alkaline phosphatase activity was also reduced with PEG grafting and metal ion release to BP. The extractable AP had shown peak activity at 72 h [for GATBP--250.5 +/- 1.2 nm pnp/mg protein/min enzyme activity (unit), PEG-GABP--165.2 +/- 16.6 units], but markedly reduced after 21 days (22.1 +/- 1.8 and 12.0 +/- 1.5 units, respectively). The initial high levels may be due to tissue injury via surgery, which mitigated with time. It is assumed that ferric ions may slow down or retard the calcification process by the inhibition of proper formation of hydroxy apatite while magnesium ions disrupt the growth of these crystals by replacing Ca2+. In addition it maybe hypothesized that these metal ions may inhibit the key element alkaline phosphatase, which acts as the substrate for mineralization. Hence, it is conceivable that a combination therapy via surface grafting of PEG and local delivery of low levels of ferric and magnesium ions may prevent the bioprosthesis associated calcification.

  12. Comparison of PTFE, pericardium bovine and fascia lata for repair of incisional hernia in rat model, experimental study.

    PubMed

    Kapan, S; Kapan, M; Goksoy, E; Karabicak, I; Oktar, H

    2003-03-01

    Incisional hernia is a frequent complication of abdominal surgery developing in 11-20 % of patients undergoing an abdominal operation. Regarding morbidity and loss of manpower, incisional hernias continue to be a fundamental problem for surgeons. In this experimental study, three commonly used mesh materials (Goretex PTFE; Tutoplast Fascia lata; Tutopatch Pericardium bovine) were compared according to effectiveness, strength, adhesion formation, histological changes, and early complications. Three groups, each consisting of 14 rats, have been formed as group A: polytetrafluoroethylene (PTFE), group B: pericardium bovine and group C: fascia lata. Evaluations were achieved at the end of the first and second postoperative week, respectively. Adhesion formation, wound maturation, bursting pressure, and tensile strength were evaluated. No statistically significant difference regarding adhesion formation was observed between groups although adhesion formation was less significant in PTFE and pericardium bovine groups than in the fascia lata group. Bursting pressure and tensile strength values were significantly higher in PTFE group than in the fascia lata group ( P<0.05). No statistically significant difference was observed between groups regarding wound maturation. In this experimental model, PTFE and pericardium bovine were found to be superior to fascia lata in abdominal wall repair.

  13. Poisson's ratio of collagen fibrils measured by small angle X-ray scattering of strained bovine pericardium

    NASA Astrophysics Data System (ADS)

    Wells, Hannah C.; Sizeland, Katie H.; Kayed, Hanan R.; Kirby, Nigel; Hawley, Adrian; Mudie, Stephen T.; Haverkamp, Richard G.

    2015-01-01

    Type I collagen is the main structural component of skin, tendons, and skin products, such as leather. Understanding the mechanical performance of collagen fibrils is important for understanding the mechanical performance of the tissues that they make up, while the mechanical properties of bulk tissue are well characterized, less is known about the mechanical behavior of individual collagen fibrils. In this study, bovine pericardium is subjected to strain while small angle X-ray scattering (SAXS) patterns are recorded using synchrotron radiation. The change in d-spacing, which is a measure of fibril extension, and the change in fibril diameter are determined from SAXS. The tissue is strained 0.25 (25%) with a corresponding strain in the collagen fibrils of 0.045 observed. The ratio of collagen fibril width contraction to length extension, or the Poisson's ratio, is 2.1 ± 0.7 for a tissue strain from 0 to 0.25. This Poisson's ratio indicates that the volume of individual collagen fibrils decreases with increasing strain, which is quite unlike most engineering materials. This high Poisson's ratio of individual fibrils may contribute to high Poisson's ratio observed for tissues, contributing to some of the remarkable properties of collagen-based materials.

  14. Poisson's ratio of collagen fibrils measured by small angle X-ray scattering of strained bovine pericardium

    SciTech Connect

    Wells, Hannah C.; Sizeland, Katie H.; Kayed, Hanan R.; Haverkamp, Richard G.; Kirby, Nigel; Hawley, Adrian; Mudie, Stephen T.

    2015-01-28

    Type I collagen is the main structural component of skin, tendons, and skin products, such as leather. Understanding the mechanical performance of collagen fibrils is important for understanding the mechanical performance of the tissues that they make up, while the mechanical properties of bulk tissue are well characterized, less is known about the mechanical behavior of individual collagen fibrils. In this study, bovine pericardium is subjected to strain while small angle X-ray scattering (SAXS) patterns are recorded using synchrotron radiation. The change in d-spacing, which is a measure of fibril extension, and the change in fibril diameter are determined from SAXS. The tissue is strained 0.25 (25%) with a corresponding strain in the collagen fibrils of 0.045 observed. The ratio of collagen fibril width contraction to length extension, or the Poisson's ratio, is 2.1 ± 0.7 for a tissue strain from 0 to 0.25. This Poisson's ratio indicates that the volume of individual collagen fibrils decreases with increasing strain, which is quite unlike most engineering materials. This high Poisson's ratio of individual fibrils may contribute to high Poisson's ratio observed for tissues, contributing to some of the remarkable properties of collagen-based materials.

  15. Satisfaction of patients with Peyronie's disease after plaque surgery and bovine pericardium graft.

    PubMed

    Silva-Garretón, A; Santillán, D; Chávez, D; Gioielli, A; Rey-Valzacchi, G; Layús, O; Gueglio, G

    2017-03-01

    Plaque and bovine pericardium graft surgery is a useful tool for treating Peyronie's disease. To determine patient satisfaction following this operation. This was a retrospective, observational and descriptive study. We collected data from the medical records of patients who underwent surgery between 2004 and 2015 and were evaluated through a postoperative satisfaction questionnaire. Twenty-eight operations were performed. Curve correction was achieved in 26 patients (95.3%). One patient (3.57%) required residual curve correction using Yachia's technique, and 1 patient (3.57%) had a severe complication consisting of prosthetic infection and urethrocutaneous fistulae. Twenty-one patients (75%) expressed satisfaction with the surgery. Our results show an acceptable level of satisfaction among our patients, with a low number of complications. However, further prospective, controlled and randomised studies are needed. Copyright © 2016 AEU. Publicado por Elsevier España, S.L.U. All rights reserved.

  16. Bovine pericardium for portal vein reconstruction in abdominal surgery: a surgical guide and first experiences in a single center.

    PubMed

    Jara, Maximilian; Malinowski, Maciej; Bahra, Marcus; Stockmannn, Martin; Schulz, Antje; Pratschke, Johann; Puhl, Gero

    2015-01-01

    Resection and reconstruction of infiltrated vessels achieve resectability of extended pancreatic tumors. The aim of the present study was to assess the feasibility of bovine pericardium as graft material for the individualised portal vein reconstruction and demonstrate a surgical technique for abdominal vein repair. We performed a MEDLINE search to review the methods for complex abdominal vein reconstruction in the course of extended pancreatectomy. Moreover, clinical data of patients receiving portal vein reconstruction using a bovine pericardial patch at our institution were retrospectively analyzed. Based on the results of a review of the literature, autologous venous grafts using the internal jugular vein represent the most popular option for segmental portal vein reconstruction in case of impossible direct suture. At our center, segmental portal vein reconstruction with bovine pericardial patch in course of pancreatic surgery was performed in 4 patients. No case of vascular complications such as occlusion, segmental stenosis or thrombosis occurred. Our experience suggests a surgical procedure for an individual size-matched portal vein reconstruction using bovine pericardium. Although first results appear promising, prospective studies are required to objectively assess the patency of bovine pericardium compared with autologous and synthetic interposition grafts for portal vein reconstruction. © 2015 S. Karger AG, Basel.

  17. Activity of MMP-9 after repair of abdominal wall defects with acellular and crosslinked bovine pericardium in rabbit.

    PubMed

    Singh, Himani; Kumar, Naveen; Sharma, A K; Kataria, Meena; Munjal, Ashok; Kumar, Amit; Dewangan, Rukmani; Kumar, Vineet; Devarathnam, J; Kumar, Sachin

    2014-02-01

    This study was undertaken for the identification of matrix metalloproteinases (MMPs) in extracts obtained from native, acellular and crosslinked bovine pericardium (in vitro), as well as in the plasma after implantation of these biomaterials in rabbits (in vivo). Native pericardium (NP) expressed a 72 kDa (MMP-2) band; whereas, in acellular pericardium (AP) two bands (10 kDa and 92 kDa) of MMPs were observed of which, 92 kDa band was very faint. AP crosslinked with glutaraldehyde did not show any gelatinase activity and thus reflects the creation of new additional chemical bonds between the collagen molecules which has been effectively removed. Gelatin zymography showed only one major band of 92 kDa in all the implanted and untreated rabbit plasma, but the relative amount of 92 kDa was 1-2 times higher in acellular bovine pericardium implanted rabbits as compared to crosslinked and native groups. In NP group, the 92 kDa band was the dullest among the three groups. This indicated that the level of MMP-9 corresponds to the degree of collagen degradation. © 2012 The Authors. International Wound Journal © 2012 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  18. Immune response to bovine pericardium implanted into α1,3-galactosyltransferase knockout mice: feasibility as an animal model for testing efficacy of anticalcification treatments of xenografts.

    PubMed

    Lee, Cheul; Ahn, Hyuk; Kim, Soo Hwan; Choi, Sun Young; Kim, Yong Jin

    2012-07-01

    Glutaraldehyde (GA)-fixed xenografts are prone to calcification after implantation in humans and there is evidence that immune reaction to the Galα1,3-Galβ1,4GlcNAc-R (α-Gal) antigen may play a part in this process. The objectives of this study were to evaluate the immune response of α1,3-galactosyltransferase knockout (α-Gal KO) mice to bovine pericardium and to evaluate the effect of various anticalcification treatments on bovine pericardium using mouse subcutaneous implantation model. Bovine pericardial tissues were divided into eight groups according to the method of anticalcification treatments. Prepared tissues were subcutaneously implanted into the α-Gal KO and wild-type mice for 2 months, and anti-α-Gal antibodies were measured at 2 weeks and 2 months after implantation. Explanted tissues were examined by immunohistochemistry and calcium contents of the explanted tissues were measured. Titres of IgM and IgG antibodies in the α-Gal KO mice increased significantly according to the duration of implantation, whereas titres of IgM and IgG antibodies in the wild-type mice increased until 2 weeks after implantation without further increase thereafter. Titres of IgG antibodies measured at 2 months after implantation were significantly higher in the α-Gal KO mice than in the wild-type mice. Immunohistochemistry revealed macrophages surrounding the pericardial tissues irrespective of the mouse type into which the tissues implanted, whereas T-cells could only be observed in the tissues implanted into the α-Gal KO mice. Except the high-concentration GA-treated group, calcium contents of anticalcification-treated groups were all significantly lower or tended to be lower than that of the control group, irrespective of the mouse type. Calcium contents of the control group were significantly higher in the α-Gal KO mice than in the wild-type mice. Bovine pericardium implanted into the α-Gal KO mice caused significant increase in anti-α-Gal antibodies, showed

  19. A prospective, multicenter study of bovine pericardium membrane with cancellous particulate allograft for localized alveolar ridge augmentation.

    PubMed

    Sterio, Thomas W; Katancik, James A; Blanchard, Steven B; Xenoudi, Pinelopi; Mealey, Brian L

    2013-01-01

    Resorption of the alveolar ridge may lead to ridge deformities that make dental implant placement difficult or impossible. Augmentation of the alveolar ridge may restore appropriate ridge form to allow implant placement. Forty-four patients with edentulous spaces completed this multicenter prospective trial to clinically and radiographically evaluate the efficacy of a bovine pericardium membrane and a particulate mineralized cancellous bone allograft in promoting lateral ridge augmentation. Overall, 38 of 44 patients (86.4%) were able to receive dental implants in the appropriate restoratively driven position 6 months after ridge augmentation. The mean gain in clinical ridge width after augmentation was 2.61 mm, while radiographically the mean gain in ridge width was 1.65 mm at a level 3 mm apical to the bony crest and 1.93 mm at a level 6 mm apical to the crest. On average, approximately 50% of the graft material added horizontally during surgery was displaced or resorbed during healing. Histomorphometric evaluation of cores taken from the augmented ridge at 6 months revealed that approximately 58% of the tissue volume was vital bone, with 12% residual allograft particles and 30% nonmineralized tissue.

  20. Guided bone regeneration using acellular bovine pericardium in a rabbit mandibular model: in-vitro and in-vivo studies.

    PubMed

    Bai, M; Zhang, T; Ling, T; Zhou, Z; Xie, H; Zhang, W; Hu, G; Jiang, C; Li, M; Feng, B; Wu, H

    2014-08-01

    To investigate the feasibility of acellular bovine pericardium (BP) for guided bone regeneration (GBR) in vitro and in vivo. The success of GBR relies on the fact that various cellular components possess different migration rates into the defect site and that a barrier membrane plays a significant role in these processes. BP membrane was isolated and decellularized using an enzymatic method. The microarchitecture, mechanical properties, cytotoxicity and cell chemotaxis properties of the acellular BP were evaluated in vitro, and the in-vivo efficacy of the acellular BP was also investigated in a rabbit mandibular model. The acellular BP membrane possessed an interconnected fibrous structure. Glutaraldehyde (GA) treatment was efficient for enhancement of the mechanical properties of the acellular BP bur and resulted in negligible cytotoxicity. After 16 wk, standardized osseous defects created in the rabbit mandible, and covered with acellular BP, were associated with an enhanced deposition of mineralized tissue when compared with defects left to spontaneous healing. GA-treated acellular BP is promising as a barrier membrane for GBR for further in-vivo and clinical studies. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  1. Usefulness of bovine pericardium as interpositional graft in the surgical repair of nasal septal perforations (experimental study).

    PubMed

    Jasso-Victoria, Rogelio; Olmos-Zuñiga, J Raul; Gutierrez-Marcos, L Miguel; Sotres-Vega, Avelina; Manjarrez Velazquez, J Ramon; Gaxiola-Gaxiola, Miguel; Avila-Chavez, Arturo; Avendaño Moreno, Guillermo; Santillan-Doherty, Patricio

    2003-01-01

    A 2.5-cm nasal septal perforation was performed in 18 pigs and repaired as follows: group I (n = 6), septal perforation without treatment; group II (n = 6), surgical repair with interpositional graft of glutaraldehyde-preserved bovine pericardium (GPBP); group III (n = 6), surgical repair with interpositional graft of lyophilized GPBP (LGPBP). The animals were evaluated clinically and radiologically (x-ray and CT scan) 2 days before surgery, daily during the first postoperative week, and weekly during the next 6 months. At the end of the study the animals were euthanized with an overdose of pentobarbital. Macroscopic and microscopic examination of the grafts and nasal septum was performed. All the animals survived the surgical procedure. Five pigs in group I showed persistence of the septal perforation. All the animals in groups II and III showed total closure of the septal perforation, with the presence of fibrotic tissue on the pericardial grafts as well as in the septal cartilage, and overall good healing. In conclusion, GPBP and LGPBP are adequate materials that can be used as interpositional grafts in the surgical closure of septal perforations in pigs

  2. Magnesium Presence Prevents Removal of Antigenic Nuclear-Associated Proteins from Bovine Pericardium for Heart Valve Engineering.

    PubMed

    Dalgliesh, Ailsa J; Liu, Zhi Zhao; Griffiths, Leigh G

    2017-03-10

    Current heart valve prostheses are associated with significant complications, including aggressive immune response, limited valve life expectancy, and inability to grow in juvenile patients. Animal derived "tissue" valves undergo glutaraldehyde fixation to mask tissue antigenicity; however, chronic immunological responses and associated calcification still commonly occur. A heart valve formed from an unfixed bovine pericardium (BP) extracellular matrix (ECM) scaffold, in which antigenic burden has been eliminated or significantly reduced, has potential to overcome deficiencies of current bioprostheses. Decellularization and antigen removal methods frequently use sequential solutions extrapolated from analytical chemistry approaches to promote solubility and removal of tissue components from resultant ECM scaffolds. However, the extent to which such prefractionation strategies may inhibit removal of antigenic tissue components has not been explored. We hypothesize that presence of magnesium in prefractionation steps causes DNA precipitation and reduces removal of nuclear-associated antigenic proteins. Keeping all variables consistent bar the addition or absence of magnesium (2 mM magnesium chloride hexahydrate), residual BP ECM scaffold antigenicity and removed antigenicity were assessed, along with residual and removed DNA content, ECM morphology, scaffold composition, and recellularization potential. Furthermore, we used proteomic methods to determine the mechanism by which magnesium presence or absence affects scaffold residual antigenicity. This study demonstrates that absence of magnesium from antigen removal solutions enhances solubility and subsequent removal of antigenic nuclear-associated proteins from BP. We therefore conclude that the primary mechanism of action for magnesium removal during antigen removal processes is avoidance of DNA precipitation, facilitating solubilization and removal of nuclear-associated antigenic proteins. Future studies are

  3. Aortic valve leaflet replacement with bovine pericardium to preserve native dynamic capabilities of the aortic annulus.

    PubMed

    Kim, Kyung Hwa; Choi, Jong Bum; Kim, Min Ho; Kim, Won Ho; Lee, Mi Kyung; Lee, Sam Youn

    2014-02-01

    Valve replacement is typically the most appropriate option for treating aortic valve stenotic insufficiency. However, neither mechanical nor bioprosthetic replacement components preserve the circumferential expansion and contraction of a native aortic annulus during the cardiac cycle, because the prosthetic ring is affixed to the annulus. A 64-year-old man presented with a bicuspid and stenotic aortic valve, and the native annulus was too small to accommodate a porcine replacement valve. We fashioned new aortic leaflets from bovine pericardium with use of a template, and we affixed the sinotubular junction with use of inner and outer stabilization rings. Postoperative echocardiograms revealed coaptation of the 3 new leaflets with no regurgitation. At the patient's 5.5-year follow-up examination, echocardiograms showed flexible leaflet movement with a coaptation height of 7 mm, and expansion and contraction of the aortic annulus similar to that of a normal native annulus. The transvalvular pressure gradient was insignificant. If long-term durability of the new leaflets is confirmed, this method of leaflet replacement and fixation of the sinotubular junction might serve as an acceptable alternative to valve replacement in the treatment of aortic valve stenosis. We describe the patient's case and present our methods and observations.

  4. Aortic Valve Leaflet Replacement with Bovine Pericardium to Preserve Native Dynamic Capabilities of the Aortic Annulus

    PubMed Central

    Kim, Kyung Hwa; Kim, Min Ho; Kim, Won Ho; Lee, Mi Kyung; Lee, Sam Youn

    2014-01-01

    Valve replacement is typically the most appropriate option for treating aortic valve stenotic insufficiency. However, neither mechanical nor bioprosthetic replacement components preserve the circumferential expansion and contraction of a native aortic annulus during the cardiac cycle, because the prosthetic ring is affixed to the annulus. A 64-year-old man presented with a bicuspid and stenotic aortic valve, and the native annulus was too small to accommodate a porcine replacement valve. We fashioned new aortic leaflets from bovine pericardium with use of a template, and we affixed the sinotubular junction with use of inner and outer stabilization rings. Postoperative echocardiograms revealed coaptation of the 3 new leaflets with no regurgitation. At the patient's 5.5-year follow-up examination, echocardiograms showed flexible leaflet movement with a coaptation height of 7 mm, and expansion and contraction of the aortic annulus similar to that of a normal native annulus. The transvalvular pressure gradient was insignificant. If long-term durability of the new leaflets is confirmed, this method of leaflet replacement and fixation of the sinotubular junction might serve as an acceptable alternative to valve replacement in the treatment of aortic valve stenosis. We describe the patient's case and present our methods and observations. PMID:24512414

  5. Effect of SDS-based decelullarization in the prevention of calcification in glutaraldehyde-preserved bovine pericardium: study in rats.

    PubMed

    Collatusso, Claudinei; Roderjan, João Gabriel; Vieira, Eduardo Discher; Costa, Francisco Diniz Affonso da; Noronha, Lucia de; Fornazari, Daniele de Fátima

    2012-01-01

    The aim of study was to investigate the SDS-based decellularization process as an anticalcification method in glutaraldehyde-preserved bovine pericardium in subcutaneous rat model. Pericardium samples with 0.5 cm² area were divide in four groups: group GDA: 0.5% glutaraldehyde-preserved pericardium (GDA); group GDA-GL: GDA + 0.2% glutamic acid (GL); group D-GDA: decellularized (D) pericardium with 0.1% SDS + GDA and group D-GDA-GL: decellularized pericardium + GDA + 0.2% glutamic acid. After this samples were implanted in 18 rats in subcutaneous position till 90 days. Each animal received samples of the four groups. The explants were performed at 45 and 90 days. The explants were subjected to histology in glass slides stained with hematoxilin-eosin and alizarin red, morphometry evaluation and the calcium content was measured by flame atomic absorption spectrometry. The inflammatory infiltrate was the same in all groups, however more intense in GDA and GDA-GL groups in 45 days, increasing at 90 days. The calcium contents for 45 days were: 32.52 ± 3.19 µg/mg in GDA group; 22.12 ± 3.87 µg/ mg in GDA-GL group; 1.06 ± 0.38 µg/mg in D-GDA group and 3.99 ± 5.78 µg/mg in D-GDA-GL (P< 0.001). For 90 days were 65.91 ± 24.67 µg/mg in GDA group; 38.37 ± 13.79 µg/mg in GDA-GL group; 1.24 ± 0.99 µg/mg in D-GDA group and 30.54 ± 8.21 µg/mg in D-GDA-GL (P< 0.001). Only D-GDA did not show increase rates of calcium at 45 to 90 days (P=0.314). SDS-based decellularization process reduced the inflammatory intensity and calcification in bovine pericardium in subcutaneous rat model for 90 days.

  6. Identification of specific calcium-binding noncollagenous proteins associated with glutaraldehyde-preserved bovine pericardium in the rat subdermal model.

    PubMed

    Gura, T A; Wright, K L; Veis, A; Webb, C L

    1997-06-15

    Calcification of glutaraldehyde-preserved bioprosthetic heart valves (BHVs) results in their clinical failure. The mechanism of this pathologic calcification is not well defined. Since serum proteins are known to be taken up in mineralized tissue, we hypothesized that serum proteins derived from several calcium-binding noncollagenous proteins (NCPs) of bone and teeth also may be associated with pathologically mineralized BHVs. Using a rat subdermal model of BHV calcification, glutaraldehyde-preserved bovine pericardium (GPBP) was implanted for 1, 3, 14, and 60 days, and then subjected to an extraction procedure designed to isolate only NCPs tightly bound to the mineral phase. Gel electrophoresis and Coomassie Brilliant Blue staining demonstrated that these proteins became associated with GPBP over time, paralleling reported calcium uptake by the tissue. Stains-All staining demonstrated a marked accumulation of highly acidic, phosphorylated NCPs associated with 60-day GPBP extracts. Some of these proteins were detected in rat serum but were absent from extracts of GPBP incubated in rat serum in vitro. Western blotting with antibodies to three NCPs found in bone and teeth-bone acidic glycoprotein 75 (BAG 75), osteopontin, and SPARC-demonstrated that these NCPs were tightly bound to the mineral phase of calcified GPBP. A fourth NCP, bone sialoprotein II (BSP II) was barely detectable. Thus each identified NCP showed a different pattern of GPBP association relative to mineral deposition, suggesting unique roles for each in pathologic calcification. SPARC increased within 3 days of GPBP implantation but decreased by 2 weeks. BAG 75 and osteopontin uptake was detected in the initial mineral deposits and increased mineralization proceeded. BSP II never increased significantly over the entire-period. Further studies, which should include immunohistochemistry, will be important for delineating the source, location, and function of these three NCPs and for identifying others

  7. Tendon elongation with bovine pericardium (Tutopatch®) when conventional strabismus surgery is not possible.

    PubMed

    van Rijn, Laurentius J; van De Ven, Stephanie J N; Krijnen, Jacqueline S M; Jansen, Suzanne M; Bakels, Amber J; Langenhorst, Anne-Marie E L

    2016-01-01

    Sometimes, a conventional recess-resect surgery may not be sufficient to obtain satisfactory ocular alignment. Patients who have previously undergone surgery and/or have a large difference in visual acuity between both eyes and do not wish to undergo surgery on the sound eye provide a surgical challenge. In these cases, tendon elongation with bovine pericardium may be an option. We retrospectively reviewed the charts of 38 patients who underwent strabismus surgery with tendon elongation. Before surgery, 31 had exotropia (angle -21.8 ± 5.7 degrees) and 7 esotropia (angle +19.1 ± 5.4 degrees). Reasons for tendon elongation included the following: 15 patients refused surgery on their sound eye; in 15 patients, conventional recess-resect was not possible; and in 7 patients, the elongation best fitted the motility pattern. In one patient, tendon elongation was preferred over conventional recession because of a thin sclera. Follow-up was 0.5 to 4 years. At last follow-up visit, in patients with previous exotropia, the angle was -3.3 ± 5.9 degrees; in patients with previous esotropia, +0.2 ± 0.5 degrees. Most had some duction limitation in the direction of the elongated muscle. All patients but one were satisfied with the result. In the patients with previous exotropia, there was a small but nonsignificant regression to recurrence of the exodeviation (on average 0.5 degree per year). Tendon elongation is a valuable addition to our strabismus surgery repertoire. However, because of duction limitations after surgery, it should be reserved for those cases in which conventional surgery is not an option.

  8. Modifications on collagen structures promoted by 1,4-dioxane improve thermal and biological properties of bovine pericardium as a biomaterial.

    PubMed

    Forti, Fábio L; Goissis, Gilberto; Plepis, Ana M G

    2006-01-01

    Collagen is a widely used raw material for biomaterial manufacture, which generally depends on chemical modifications of this fibrillar protein with cross-linking agents to improve biocompatibility and mechanical properties. However, cross-linking reduces the natural properties of collagen, such as low immune response, low toxicity as well as the ability to promote cellular growth and attachment. In this work, the modifications promoted by 1,4-dioxane solvent on the collagen present in native bovine pericardium (NBP) matrix routinely used in bioprosthesis manufacture, with or without subsequent cross-linking by glutaraldehyde, has been studied. The structural changes of NBP evaluated by scanning electron microscopy show that 1,4-dioxane induces a more homogeneous material by increasing aggregation of collagen fibers, while transmission scanning electron microscopy shows that natural collagen fibril arrangement, integrity, and the D-periodicity pattern are maintained by solvent treatments. Measurements of thermal stability and resistance to collagenase enzymatic digestion of NBP matrices treated with 1,4-dioxane show an increase in melting temperature and decrease in biodegradability, as compared to native pericardium. Cross-linking with glutaraldehyde improves all the analyzed NBP properties, which are not impaired by previous treatment with 1,4-dioxane. Histological evaluation of NBP submitted to 1,4-dioxane treatment shows lower lipid and cell contents and improvement in other morphologic characteristics compared to native pericardium. Altogether, these results suggest the use of 1,4-dioxane organic solvent as an alternative non-cross-linking treatment for direct utilization on rich collagen matrices, resulting in materials with improved biocompatibility and physicochemical properties suitable for tissue engineering.

  9. In vivo effects of human adipose-derived stem cells reseeding on acellular bovine pericardium in nude mice

    PubMed Central

    Dai, Miao; Xu, Peirong; Hou, Min; Teng, Yincheng; Feng, Jie

    2015-01-01

    Tissue-engineered biologic products may be a viable option in the reconstruction of pelvic organ prolapse (POP). This study was based on the hypothesis that human adipose-derived stem cells (hASCs) are viable in acellular bovine pericardium (ABP), when reseeded by two different techniques, and thus, aid in the reconstruction. To investigate the reseeding of hASCs on ABP grafts by using non-invasive bioluminescence imaging (BLI), and to identify the effective hASCs–scaffold combinations that enabled regeneration. Thirty female athymic nude mice were randomly divided into three groups: In the VIVO group, ABPs were implanted in the subcutaneous pockets and enhanced green fluorescent protein luciferase (eGFP·Luc)-hASCs (1 × 106 cells/50 µL) were injected on the ABP at the same time. In the VITRO group, the mice were implanted with grafts that ABP were co-cultured with eGFP·Luc-hASCs in vitro. The BLANK group mice were implanted with ABP only. The eGFP·Luc-hASCs reseeded on ABP were analyzed by BLI, histology, and immunohistochemistry. The eGFP·Luc-hASCs reseeded on ABP could be visualized at 12 weeks in vivo. Histology revealed that the VIVO group displayed the highest cell ingrowths, small vessels, and percent of collagen content per unit area. Desmin and α-smooth muscle actin were positive at the same site in the VIVO group cells. However, few smooth muscles were observed in the VITRO and BLANK groups. These results suggest that hASCs reseeded on ABP in vivo during surgery may further enhance the properties of ABP and may promote regeneration at the recipient site, resulting in a promising treatment option for POP. PMID:26253192

  10. In vivo effects of human adipose-derived stem cells reseeding on acellular bovine pericardium in nude mice.

    PubMed

    Wu, Qingkai; Dai, Miao; Xu, Peirong; Hou, Min; Teng, Yincheng; Feng, Jie

    2016-01-01

    Tissue-engineered biologic products may be a viable option in the reconstruction of pelvic organ prolapse (POP). This study was based on the hypothesis that human adipose-derived stem cells (hASCs) are viable in acellular bovine pericardium (ABP), when reseeded by two different techniques, and thus, aid in the reconstruction. To investigate the reseeding of hASCs on ABP grafts by using non-invasive bioluminescence imaging (BLI), and to identify the effective hASCs-scaffold combinations that enabled regeneration. Thirty female athymic nude mice were randomly divided into three groups: In the VIVO group, ABPs were implanted in the subcutaneous pockets and enhanced green fluorescent protein luciferase (eGFP·Luc)-hASCs (1 × 10(6) cells/50 µL) were injected on the ABP at the same time. In the VITRO group, the mice were implanted with grafts that ABP were co-cultured with eGFP·Luc-hASCs in vitro. The BLANK group mice were implanted with ABP only. The eGFP·Luc-hASCs reseeded on ABP were analyzed by BLI, histology, and immunohistochemistry. The eGFP·Luc-hASCs reseeded on ABP could be visualized at 12 weeks in vivo. Histology revealed that the VIVO group displayed the highest cell ingrowths, small vessels, and percent of collagen content per unit area. Desmin and α-smooth muscle actin were positive at the same site in the VIVO group cells. However, few smooth muscles were observed in the VITRO and BLANK groups. These results suggest that hASCs reseeded on ABP in vivo during surgery may further enhance the properties of ABP and may promote regeneration at the recipient site, resulting in a promising treatment option for POP. © 2016 by the Society for Experimental Biology and Medicine.

  11. Use of Bovine Pericardium and Sutureless Biological Glue in Left Ventricular Rupture After Mitral Valve Replacement, Five Years of Follow-up.

    PubMed

    Costa, Mário Augusto Cray da; Laforga, Fernando Cesar; Maftum, Josué Abrão; Favaro, Mateus Gustavo

    2015-01-01

    Rupture of the left ventricular wall after mitral valve replacement is an infrequent but lethal complication. Reporting correction technique of ventricular rupture with bovine pericardium patch secured with glue and without suturing: a 51 years-old female patient, with double rheumatic mitral lesion, severe stenosis and discrete insufficiency, who had a mitral valve replacement. During surgery, the patient presented a ventricular rupture of the posterior wall (atrioventricular disruption), which was successfully repaired using bovine pericardium with sutureless biological glue over the epicardium of the damaged area. Sixty months after surgery the patient has no symptoms.

  12. Evaluation of the anti-migration effect of barbed prostatic stents: in vitro study in urethra-mimicking bovine pericardium phantoms.

    PubMed

    Na, Han Kyu; Song, Ho-Young; Kim, Jin Hyoung; Nam, Deok Ho; Park, Jung-Hoon; Jeong, In Gab; Kim, Choung-Soo

    2013-02-01

    This study was designed to compare the migration rates of covered stents with eight, four, and zero barbs in urethra-mimicking bovine pericardium models. Three types of polytetrafluoroethylene-covered retrievable self-expandable prostatic stents, with eight, four, and zero barbs, were tested in bovine pericardium models, consisting of normal saline-filled acrylic containers with tubes at both ends and a replica of the human urethra. The barbs were 2 mm in length and projected 60° toward the urinary bladder. The anti-migration force (AMF) and resistance force against stent removal (RFSR) were measured by an automatic push-pull gauge system at a fixed rate (2 mm/s). Significant differences in AMF among the three stent types were detected (P < 0.001). Median AMF was far greater for eight-barbed than for four-barbed stents (11.96 vs. 7.55 N, P = 0.003). For stent removal, the difference between 4- and 8-barbed stents were not significant in median static (5.54 vs. 6.08 N, P = 0.15) or sliding (prostatic urethra, 5.32 vs. 5.59 N, P = 0.65; membranous urethra, 7.78 vs. 8.57 N, P = 0.364) RFSR. No perforations or scratched traces were observed during removal of these stents. The inclusion of eight barbs projecting 60° toward the urinary bladder were not only effective against stent migration in this model but suggests that these stents may be suitable for the treatment of prostatic urethral strictures without severely irritating the tissue. However, whether these results translate into living urethral tissue is unclear, necessitating further studies.

  13. Feasibility study of collagen membranes derived from bovine pericardium and intestinal serosa for the repair of cranial defects in ovariectomised rats.

    PubMed

    Hirata, H H; Munhoz, M A S; Plepis, A M G; Martins, V C A; Santos, G R; Galdeano, E A; Cunha, M R

    2015-07-01

    The indication of biomaterials has increased substantially in the regenerative therapy of bone defects. However, in addition to evaluating the physicochemical properties of biomaterials, the quality of the recipient tissue is also essential for the osseointegration of implants, as abnormalities in bone metabolism, such as gonadal hormone deficiency, can influence bone healing. This study evaluated the osteoregenerative capacity of collagen membranes derived from bovine pericardium and intestinal serosa in the repair of cranial defects in ovariectomised rats. Thirty female Wistar rats were submitted to surgical creation of a 5-mm cranial bone defect. The rats were divided into a control group (not ovariectomised) and an ovariectomised group. The non-ovariectomised group was divided into three subgroups: control (G1) in which the defect was not filled with the biomaterial, and two subgroups (G2 and G3) that received the bovine pericardium- and serosa-derived collagen membranes, respectively. The ovariectomised group was divided into the same subgroups (G4, G5, and G6). The animals were sacrificed 8 weeks after surgery. The calvaria were removed for macroscopic and radiographic photodocumentation and processed for histomorphometric analysis of bone healing at the surgical site. Macroscopic, radiological, and microscopic analyses demonstrated the biocompatibility of the implanted collagen membranes, as indicated by the absence of infiltration and signs of inflammation at the surgical site. Histologically, discrete immature bone neoformation projecting from the margins of the defect was observed at the surgical site in ovariectomised groups when compared to the non-ovariectomised groups. The volume of newly formed bone was significantly higher in the non-ovariectomised groups (G1: 7.83%±1.32; G2: 21.33%±1.96; and G3: 22.83%±0.98) compared to the respective ovariectomised subgroups (G4: 3.16%±0.75; G5: 16.83%±0.98; and G6: 16.16%±0.75), thus demonstrating the

  14. Triglycidylamine Crosslinking of Porcine Aortic Valve Cusps or Bovine Pericardium Results in Improved Biocompatibility, Biomechanics, and Calcification Resistance

    PubMed Central

    Connolly, Jeanne M.; Alferiev, Ivan; Clark-Gruel, Jocelyn N.; Eidelman, Naomi; Sacks, Michael; Palmatory, Elizabeth; Kronsteiner, Allyson; DeFelice, Suzanne; Xu, Jie; Ohri, Rachit; Narula, Navneet; Vyavahare, Narendra; Levy, Robert J.

    2005-01-01

    We investigated a novel polyepoxide crosslinker that was hypothesized to confer both material stabilization and calcification resistance when used to prepare bioprosthetic heart valves. Triglycidylamine (TGA) was synthesized via reacting epichlorhydrin and NH3. TGA was used to crosslink porcine aortic cusps, bovine pericardium, and type I collagen. Control materials were crosslinked with glutaraldehyde (Glut). TGA-pretreated materials had shrink temperatures comparable to Glut fixation. However, TGA crosslinking conferred significantly greater collagenase resistance than Glut pretreatment, and significantly improved biomechanical compliance. Sheep aortic valve interstitial cells grown on TGA-pretreated collagen did not calcify, whereas sheep aortic valve interstitial cells grown on control substrates calcified extensively. Rat subdermal implants (porcine aortic cusps/bovine pericardium) pretreated with TGA demonstrated significantly less calcification than Glut pretreated implants. Investigations of extracellular matrix proteins associated with calcification, matrix metalloproteinases (MMPs) 2 and 9, tenascin-C, and osteopontin, revealed that MMP-9 and tenascin-C demonstrated reduced expression both in vitro and in vivo with TGA crosslinking compared to controls, whereas osteopontin and MMP-2 expression were not affected. TGA pretreatment of heterograft biomaterials results in improved stability compared to Glut, confers biomechanical properties superior to Glut crosslinking, and demonstrates significant calcification resistance. PMID:15631995

  15. Analysis of shearing stress in the limited durability of bovine pericardium used as a biomaterial.

    PubMed

    Carrera San Martin, A; García Paez, J M; García Sestafe, J V; Herrero, E J; Navidad, R; Cordón, A; Castillo-Olivares, J L

    1998-02-01

    The objective of the study was to determine the shearing stress exerted by the suture thread under conditions of normal working stress. Thirty-six samples of calf pericardium, similar to that employed in the manufacture of bioprosthetic cardiac valve leaflets, were subjected to tensile testing. Prior to the trial, a continuous suture was sewn in the central zone of each sample, at a 45 degrees angle to the longest axis of the sample, using commercially-available threads (silk, Gore-Tex, Surgilene and nylon). Application of the Mohr circle for combined wear revealed that the shearing stress ranged between 2.68-fold greater (for samples sewn with silk) and 5.48-fold greater (for samples sewn with nylon) than the working tensile stress in the region of the suture. It is concluded that the shearing stress is responsible for the limited durability of sutured samples of calf pericardium prepared to simulate bioprosthetic cardiac valve leaflets.

  16. Bovine pericardium based non-cross linked collagen matrix for successful root coverage, a clinical study in human

    PubMed Central

    2012-01-01

    Introduction The aim of this study was to clinically assess the capacity of a novel bovine pericardium based, non-cross linked collagen matrix in root coverage. Methods 62 gingival recessions of Miller class I or II were treated. The matrix was adapted underneath a coronal repositioned split thickness flap. Clinical values were assessed at baseline and after six months. Results The mean recession in each patient was 2.2 mm at baseline. 6 Months after surgery 86.7% of the exposed root surfaces were covered. On average 0,3 mm of recession remained. The clinical attachment level changed from 3.5 ± 1.3 mm to 1,8 ( ± 0,7) mm during the observational time period. No statistically significant difference was found in the difference of probing depth. An increase in the width of gingiva was significant. With a baseline value of 1.5 ± 0.9 mm an improvement of 2.4 ± 0.8 mm after six month could be observed. 40 out of 62 recessions were considered a thin biotype at baseline. After 6 months all 62 sites were assessed thick. Conclusions The results demonstrate the capacity of the bovine pericardium based non-cross linked collagen matrix for successful root coverage. This material was able to enhance gingival thickness and the width of keratinized gingiva. The percentage of root coverage achieved thereby is comparable to existing techniques. This method might contribute to an increase of patient's comfort and an enhanced aesthetical outcome. PMID:22390875

  17. Evaluation of kangaroo pericardium as an alternative substitute for reconstructive cardiac surgery.

    PubMed

    Neethling, W M L; Cooper, S; Van Den Heever, J J; Hough, J; Hodge, A J

    2002-06-01

    Bioprosthetic materials (human, bovine and porcine) are used in various cardio-thoracic repair and replacement procedures because of excellent performance and low thrombogenicity. These bioprosthetic substitutes fail due to degeneration and calcification. This study examines the morphology, tensile properties and calcification potential of kangaroo pericardium in vitro and in vivo. Bovine (control tissue) and kangaroo pericardium, fixed in 0.625% buffered glutaraldehyde, were examined by light and scanning electron microscopy. A standard method was used for biaxial testing. Pericardial strips (10 x 5 mm) were implanted subcutaneously into male Wistar rats and retrieved after 4, 6 and 8 weeks and examined by Von Kossa's stain technique and atomic absorption spectrophotometry. Histology revealed serosa and fibrosa cell layers in both tissues. Electron microscopy showed a densely arranged collagen matrix in kangaroo pericardium. Kangaroo pericardium calcified significantly less than bovine pericardium at 4 weeks (0.80+/-0.28 versus 21.60+/-4.80 microg/mg) at 6 weeks (0.48+/-0.08 versus 32.80+/-14.4 microg/mg) and at 8 weeks (2.40+/-1.20 versus 30.40+/-17.20 microg/mg), respectively. Kangaroo pericardium has a densely arranged collagen matrix with a higher extensibility and significantly lower calcification potential. Therefore, kangaroo pericardium could be used as an alternative substitute in cardiac surgery because of its low calcification potential.

  18. Comparison of Lyophilized Glutaraldehyde-Preserved Bovine Pericardium with Different Vascular Prostheses for Use as Vocal Cords Implants: Experimental Study

    PubMed Central

    Olmos-Zuñiga, J. Raúl; Jasso-Victoria, Rogelio; Gaxiola-Gaxiola, Miguel; Sotres-Vega, Avelina; Hernández-Jiménez, Claudia; Baltazares-Lipp, Matilde; Arredondo del Bosque, Fernando; Santillan-Doherty, Patricio

    2015-01-01

    This study compared the use of lyophilized glutaraldehyde-preserved bovine pericardium (LGPBP), polytetrafluoroethylene (PTFE), polyethylene terephthalate (PET), and Teflon felt (TF) as implants for vocal cords (VC) medialization and aimed to assess the endoscopic, macroscopic, and microscopic VC changes after medialization in a canine model. In 18 mongrel dogs, the right VC were medialized with LGPBP and the left were implanted as follows: Group I (n = 6): LGPBP and PTFE; Group II (n = 6): LGPBP and PET; Group III (n = 6): LGPBP and TF. Surgical handling of the implants was compared. Three months after surgery, macroscopic and microscopic changes of VC and implants were evaluated. LGPBP offered the best surgical handling (p = 0.005, Kruskal-Wallis). TF implants showed extrusion (p = 0.005, Kruskal-Wallis) and severe inflammation. All VC formed fibrous capsules around the implants; the ones developed by LGPBP implants were thinner (p = 0.001, ANOVA, Tukey). VC implanted with synthetic materials showed eosinophilic infiltration (p = 0.01, Kruskal-Wallis). We concluded that the LGPBP could be used as an implant for VC medialization because it is biocompatible, easy to handle and remove during surgical procedures, and nonabsorbable or extrudable and produces an inflammatory reaction similar to PTFE and PET. PMID:26075232

  19. Evaluation of an established pericardium patch for delivery of mesenchymal stem cells to cardiac tissue.

    PubMed

    Vashi, Aditya V; White, Jacinta F; McLean, Keith M; Neethling, William M L; Rhodes, David I; Ramshaw, John A M; Werkmeister, Jerome A

    2015-06-01

    The present study has evaluated a commercial pericardial material for its capacity to assist as a natural extracellular matrix (ECM) patch for the delivery and retention of mesenchymal stem cells for cardiac repair. The repair of cardiac tissue with cells delivered by an appropriate bioscaffold is expected to offer a superior, long-lasting treatment strategy. The present material, CardioCel®, is based on acellular pericardium that has been stabilized by treatments, including a low concentration of glutaraldehyde, that eliminate calcification after implantation. In the present study, we have assessed this material using human bone marrow mesenchymal stem cells at various cell densities under standard, static cell culture conditions. The initial seeding densities were monitored to evaluate the extent of cell attachment and cell viability, with subsequent cell proliferation assessed up to 4 weeks using an MTS assay. Cell morphology, infiltration, and spreading were tracked using scanning electron microscopy and phalloidin staining. The efficacy of long-term cell survival was further assessed by examining the extent and type of new tissue formation on seeded scaffolds at 70 days; both type I and type III collagens were present in fibrillar structures on these scaffolds indicating that the seeded stem cells had the capacity to differentiate into collagen-producing cells necessary to repair damaged ECM. These data show that the CardioCel® scaffold is an appropriate substrate for the stem cells and has the potential to both retain seeded stem cells and to act as a template for cell propagation and new tissue formation. © 2014 Wiley Periodicals, Inc.

  20. Indentation hardness: A simple test that correlates with the dissipated-energy predictor for fatigue-life in bovine pericardium membranes for bioprosthetic heart valves.

    PubMed

    Tobaruela, Almudena; Rojo, Francisco Javier; García Paez, José María; Bourges, Jean Yves; Herrero, Eduardo Jorge; Millán, Isabel; Alvarez, Lourdes; Cordon, Ángeles; Guinea, Gustavo V

    2016-08-01

    The aim of this study was to evaluate the variation of hardness with fatigue in calf pericardium, a biomaterial commonly used in bioprosthetic heart valves, and its relationship with the energy dissipated during the first fatigue cycle that has been shown to be a predictor of fatigue-life (García Páez et al., 2006, 2007; Rojo et al., 2010). Fatigue tests were performed in vitro on 24 pericardium specimens cut in a root-to-apex direction. The specimens were subjected to a maximum stress of 1MPa in blocks of 10, 25, 50, 100, 250, 500, 1000 and 1500 cycles. By means of a modified Shore A hardness test procedure, the hardness of the specimen was measured before and after fatigue tests. Results showed a significant correlation of such hardness with fatigue performance and with the energy dissipated in the first cycle of fatigue, a predictor of pericardium durability. The study showed indentation hardness as a simple and reliable indicator of mechanical performance, one which could be easily implemented in improving tissue selection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Use of bovine pericardium (Tutopatch®) graft for surgical repair of deep melting corneal ulcers in dogs and corneal sequestra in cats.

    PubMed

    Dulaurent, Thomas; Azoulay, Thierry; Goulle, Frédéric; Dulaurent, Alice; Mentek, Marielle; Peiffer, Robert L; Isard, Pierre-François

    2014-03-01

    To evaluate the efficacy of bovine pericardium (BP) graft in the treatment of deep melting corneal ulcers in three dogs and corneal sequestra in three cats. Three dogs with keratomalacia affecting the deep third of the stroma and three cats with corneal sequestrum were evaluated and underwent surgery. Following keratectomy, BP material was placed into the keratectomy bed and sutured to the recipient cornea with 9/0 polyglactin suture material. Postoperative treatment with topical and systemic antibiotics, systemic nonsteroidal anti-inflammatory agents, and topical atropine was prescribed. Follow-up examinations were carried out 1, 2 weeks, 1 and 2 months after the surgery and consisted of a complete ophthalmic examination. Optical coherence tomography (OCT) was performed 1 and 2 months after the surgery in one dog and in one cat. At 1 week, corneal neovascularization was present around the BP graft in all cases. Four weeks after the BP graft, in two dogs and in all cats, the vascularization was regressing and the graft was integrated into the cornea, which was regaining transparency. Topical treatment with anti-inflammatory agents was then prescribed for 2 weeks. Two months after the surgery, 5 of 6 corneas in two dogs and three cats had healed with focal corneal scarring. The remaining dog had progression of the keratomalacia involving the deep BP graft that required additional surgery, but became blind. Bovine pericardium graft offers a promising option for surgical reconstruction of the cornea following keratectomy for the management of corneal ulcers and sequestra. © 2013 American College of Veterinary Ophthalmologists.

  2. Correlation between structural changes and acute thrombogenicity in transcatheter pericardium valves after crimping and balloon deployment.

    PubMed

    Bourget, J-M; Zegdi, R; Lin, J; Wawryko, P; Merhi, Y; Convelbo, C; Mao, J; Fu, Y; Xu, T; Merkel, N O; Wang, L; Germain, L; Zhang, Z; Guidoin, R

    2017-03-01

    Transcathether heart valve replacement has gained considerable acceptance during the last decades. It is now part of the armamentarium for aortic valve replacement. The procedure proved to be highly efficient. However the issues of the blood compatibility and tissue durability were not raised and the adverse events were probably under-reported, according to observations of thrombosis after deployment. Bovine pericardium leaflets were sewn inside a 26mm diameter stainless steel stent to manufacture these valves (one control and two experimental). The correlation between the trauma and the acute thombogenicity of bovine pericardium leaflets, after crimping and ballooning, was investigated via an in vitro blood flow with labeled platelets. These leaflets were processed for histology: scanning electron microscopy, light microscopy, and transmission electron microscopy. The control specimens showed a regular pericardium structure with some blood cells deposited on the collagen fibrous surface (inflow) and scarce blood cells deposited on the serous surface (outflow). After crimping and ballooning, the structure of the pericardium was severely injured, eventually with delaminations and ruptures. The blood cell uptake was considerably increased compared to the control. It would therefore be appropriate to pay more attention to the design of the valves. Specifically, the incorporation of a buffer tissue or fabric between the pericardium and the metallic stent is suggested. The issue of ballooning deserves detailed and in depth investigation regarding the lifetime of the device. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  3. New engineering treatment of bovine pericardium confers outstanding resistance to calcification in mitral and pulmonary implantations in a juvenile sheep model.

    PubMed

    Brizard, Christian P; Brink, Johann; Horton, Steven B; Edwards, Glenn Anthony; Galati, John C; Neethling, William M L

    2014-12-01

    To conduct a test of noninferiority for CardioCel (Admedus, Brisbane, Australia), a chemically engineered bovine pericardium over autologous pericardium treated intraoperatively with glutaraldehyde in a chronic juvenile sheep model of pulmonary valve (PV) and mitral valve (MV) reconstruction. We replaced the posterior leaflet of the MV and of 1 PV cusp with patches in ewes aged 10 months. There were 2 groups: CardioCel (n = 6) and control (n = 4). All valves were competent. Echocardiography was performed before euthanasia. The collected data were function, macroscopy, histology, and calcium contents. The primary end points were thickening and calcium content. All animals survived until sacrifice after 7 months. The valves had normal echo. The macroscopic aspect of the valves was excellent. Examination of the slides for both groups revealed a continuous endothelium on both sides of the patch and a layer of new collagen developed on both sides between patch and endothelium and interstitial cells and smooth muscle cell in these layers. The patch had not thickened but the 2 layers of new collagen for the PV showed a median thickening of 37% in the CardioCel group and 111% in the control group (P = .01), and for the MV a thickening of 108% and 251%, respectively, was seen (P = .01). The median calcium content in the PV was 0.24 μg/mg (range, 0.19-0.30) in the CardioCel group versus 0.34 μg/mg (range, 0.24-0.62) in controls (P = .20). In the MV it was 0.46 μg/mg (range, 0.30-1.0) in the CardioCel group and 0.47 μg/mg (range, 0.29-1.9) in controls (P = 1.0). In this growing lamb model the CardioCel patch allowed accurate valve repair at both systemic and pulmonary pressure. The mechanical properties of CardioCel after 7 months were preserved with a more controlled healing than the treated autologous pericardium and without calcification. Copyright © 2014 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.

  4. Histology of Pericardial Tissue Substitutes Used in Congenital Heart Surgery.

    PubMed

    Majeed, Amara; Baird, Christopher; Borisuk, Michele J; Sanders, Stephen P; Padera, Robert F

    Pericardium is used as a tissue substitute during pediatric heart surgery. However, little is known about the histological characteristics of pericardial tissue substitutes. We searched our clinical and Pathology databases to identify cases in which glutaraldehyde-preserved autologous pericardium, PhotoFix bovine pericardium, or Peri-Guard Repair Patch glutaraldehyde-preserved bovine pericardium was used as patch material during cardiac surgery and in which explanted tissue substitute was available for histologic examination. Tissue sections were stained with hematoxylin and eosin, Masson trichrome, and Movat pentachrome, and were graded for inflammation, tissue substitute degeneration, neointima formation, and calcification. Nonparametric statistical methods were used to test differences between groups because of small sample size. The eight patients who received PhotoFix pericardium were older (median 10 months vs 10 days, P < 0.05) and the material was in situ longer (median 14 vs 2.5 months, P < 0.05) compared to the eight who received autologous pericardium. Only three patients received glutaraldehyde-preserved bovine pericardium precluding statistical comparison. Inflammation and tissue degeneration were greater in PhotoFix pericardium compared to autologous pericardium but were no more than moderate. Neointima formation and calcification did not differ significantly between the two groups. PhotoFix bovine pericardium is associated with more inflammation and material degeneration but calcification, and neointima formation are similar to autologous pericardium. Although the short-term outcomes are acceptable, calcification and degeneration seen in some cases suggest that long-term outcomes and performance at certain anatomic locations need further study.

  5. Boxer's pericardium.

    PubMed

    Ooi, A; Douds, A C; Kumar, E B; Nashef, S A M

    2003-12-01

    A 65-year-old retired professional boxer presented with progressively worsening shortness of breath, peripheral oedema and mild abdominal swelling over a period of 6 months. His only past medical history was hypertension. Subsequent investigations revealed chylous ascites, pericardial constriction and bilateral chylothorax. He had uneventful pericardectomy, and post-operatively the chylothorax resolved only after administration of octreotide for 10 days. The histopathological features of fibrosis, haemosiderin deposition in the pericardium and abundant haemosiderin-laden macrophages are consistent with chronic resolving haemopericardium. These findings suggested that the cause of pericardial constriction was repeated chest trauma from boxing.

  6. Oxytocin binding sites in bovine mammary tissue

    SciTech Connect

    Zhao, Xin.

    1989-01-01

    Oxytocin binding sites were identified and characterized in bovine mammary tissue. ({sup 3}H)-oxytocin binding reached equilibrium by 50 min at 20{degree}C and by 8 hr at 4{degree}C. The half-time of displacement at 20{degree}C was approximately 1 hr. Thyrotropin releasing hormone, adrenocorticotropin, angiotensin I, angiotensin II, pentagastrin, bradykinin, xenopsin and L-valyl-histidyl-L-leucyl-L-threonyl-L-prolyl-L-valyl-L-glutamyl-L-lysine were not competitive. In the presence of 10 nM LiCl, addition of oxytocin to dispersed bovine mammary cells, in which phosphatidylinositol was pre-labelled, caused a time and dose-dependent increase in radioactive inositiol monophosphate incorporation. The possibility that there are distinct vasopressin receptors in bovine mammary tissue was investigated. ({sup 3}H)-vasopressin binding reached equilibrium by 40 min at 20{degree}. The half-time of displacement at 20{degree}C was approximately 1 hr. The ability of the peptides to inhibit ({sup 3}H)-vasopressin binding was: (Thr{sup 4},Gly{sup 7})-oxytocin > Arg{sup 8}-vasopressin > (lys{sup 8})-vasopressin > (Deamino{sup 1},D-arg{sup 8})-vasopressin > oxytocin > d (CH{sub 2}){sub 5}Tyr(Me)AVP.

  7. Inhibition of mineralization of glutaraldehyde-pretreated bovine pericardium by AlCl3. Mechanisms and comparisons with FeCl3, LaCl3, and Ga(NO3)3 in rat subdermal model studies.

    PubMed Central

    Webb, C. L.; Schoen, F. J.; Flowers, W. E.; Alfrey, A. C.; Horton, C.; Levy, R. J.

    1991-01-01

    In the present study, the authors investigated the mechanism by which Al3+ preincubations inhibited the pathologic calcification of glutaraldehyde-pretreated bovine pericardium (GPBP) implanted subdermally in rats. The concentration dependency of the Al3+ anticalcification effect was compared with that of other trivalent metal ions (Fe3+, Ga3+, La3+) known to interact with calcium phosphates. In vitro incubations of GPBP were carried out in AlCl3 (10(-3) mol/l [molar] to 10(-1) mol/l) to ascertain both the optimal conditions for uptake of Al3+ and the time course of Al3+ dissociation. Al3+ uptake by GPBP was concentration dependent and occurred rapidly, with tissue levels after 1 hour not differing significantly from those after 72 hours of incubation. Analyses of GPBP samples preincubated in AlCl3 (0.1 mol/l, 24 hours) showed that more than 75% of the Al3+ remained tightly bound after 60 days' in vitro release at 37 degrees C, pH 7.4. Preincubations of GPBP in AlCl3 significantly inhibited calcification after subdermal implantation in rats for 60 days (Ca++ = 5.1 +/- 0.9 microgram/mg, 11.5 +/- 4.6 micrograms/mg, 70.3 +/- 23.0 micrograms/mg, mean +/- standard error [SE], for 10(-1) mol/l, 10(-2) mol/l, 10(-3) mol/l AlCl3, respectively), compared with controls (Ca++ = 110.0 +/- 9.3 micrograms/mg). All animals were free of Al3(+)-mediated adverse effects on bone, as determined by light microscopic evaluation of femoral epiphyseal growth plates. Transmission electron microscopy coupled with electron energy loss spectroscopy (EELS) of GPBP incubated in 10(-1) mol l AlCl3 for 24 hours demonstrated discrete Al3+ localization in the sarcolemma and cytoplasmic and nuclear membranes of devitalized pericardial connective tissue cells at intracellular sites coincident with phosphorus loci. Similar intracellular localization remained prominent in explants removed after 60 days; no calcific deposits were noted in these specimens. Preincubations in Fe3+ but not Ga3+ and La3

  8. The fate of preserved homograft pericardium and autogenous pericardium within the heart

    PubMed Central

    Clarke, C. P.; Barratt-Boyes, B. G.; Sims, F. H.

    1968-01-01

    Animal experiments were performed to assess the usefulness of preserved homograft pericardium for intracardiac surgery, with particular reference to the problem of homograft mitral valve papillary muscle fixation. The fate of fresh autogenous pericardium was also studied. Both homograft and autogenous pericardium were destroyed and replaced by fibrous tissue. This reaction was faster when the pericardium was in direct contact with the blood stream and slower when it was buried within the myocardium. As homograft pericardium provided a more intense fibrous reaction than autogenous pericardium, it was considered to be very suitable for papillary muscle fixation and less suitable for other intracardiac procedures. The technique used for attaching the papillary muscle of a homograft mitral valve is briefly described. Images PMID:5654067

  9. Dynamic compressive response of bovine liver tissues.

    PubMed

    Pervin, Farhana; Chen, Weinong W; Weerasooriya, Tusit

    2011-01-01

    This study aims to experimentally determine the strain rate effects on the compressive stress-strain behavior of bovine liver tissues. Fresh liver tissues were used to make specimens for mechanical loading. Experiments at quasi-static strain rates were conducted at 0.01 and 0.1 s(-1). Intermediate-rate experiments were performed at 1, 10, and 100 s(-1). High strain rate (1000, 2000, and 3000 s(-1)) experiments were conducted using a Kolsky bar modified for soft material characterization. A hollow transmission bar with semi-conductor strain gages was used to sense the weak forces from the soft specimens. Quartz-crystal force transducers were used to monitor valid testing conditions on the tissue specimens. The experiment results show that the compressive stress-strain response of the liver tissue is non-linear and highly rate-sensitive, especially when the strain rate is in the Kolsky bar range. The tissue stiffens significantly with increasing strain rate. The responses from liver tissues along and perpendicular to the liver surface were consistent, indicating isotropic behavior.

  10. Current usage and future directions for the bovine pericardial patch

    PubMed Central

    Li, Xin; Guo, Yuanyuan; Ziegler, Kenneth; Model, Lynn; Eghbalieh, Sammy D. D.; Brenes, Robert; Kim, Susun; Shu, Chang; Dardik, Alan

    2010-01-01

    Bovine pericardium is widely used in surgery and is commonly used for a patch after arteriotomy during cardiovascular surgery. Bovine pericardial patches have several advantages compared to prosthetic patches, including superior biocompatability, easy handling, less suture line bleeding and possibly reduced rates of infection. These advantages of bovine pericardium have led to its common use during carotid endarterectomy. However, long-term clinical results reported after carotid endarterectomy have suggested several issues that may be related to the patch including restenosis, pseudoaneurysm formation, infection, fibrosis, calcification and thrombosis. These complications may diminish the long-term efficacy of carotid endarterectomy and suggest potential areas for improvement of surgical patches. Understanding the mechanisms by which bovine pericardium heals after patch angioplasty may lead to next generation tissue engineered patches. PMID:21276709

  11. New treatments using alginate in order to reduce the calcification of bovine bioprosthetic heart valve tissue.

    PubMed

    Shanthi, C; Rao, K P

    1997-01-01

    Calcification limits the functional lifetime of cardiac valve substitutes fabricated from glutaraldehyde preserved bovine pericardium. Host factors, mainly younger age, and implant factors, mainly glutaraldehyde cross-linking, are implicated in the calcification process. Glutaraldehyde cross-linking is believed to activate the potential sites in the tissues for biocalcification. In the present work, we investigated the possibility of using alginate azide (AA) instead of glutaraldehyde for the preservation of pericardial tissues in order to enhance the durability of bioprosthetic heart valves. Grafting with poly(GMA-BA) copolymer to the alginate azide cross-linked pericardial (AACPC) tissue was carried out to obtain better stability, strength, and anticalcification properties. The strength property and thermal stability of the AA cross-linked tissues were studied. Calcification studies in rat subdermal models reveal that AA cross-linking reduces the calcification to negligible levels. After 30 days implantation, the calcium content was found to be 10.4 +/- 1.2 and 6.1 +/- 0.3 micrograms mg-1 for untreated AACPC and polymer grafted AACPC, respectively, compared to a value of 100 +/- 1.2 micrograms mg-1 calcium recorded for control glutaraldehyde cross-linked pericardial (GCPC) tissues.

  12. Nanocoating with titanium reduces iC3b- and granulocyte-activating immune response against glutaraldehyde-fixed bovine pericardium: a new technique to improve biologic heart valve prosthesis durability?

    PubMed

    Guldner, Norbert W; Bastian, Fabienne; Weigel, Günther; Zimmermann, Hanngörg; Maleika, Markus; Scharfschwerdt, Michael; Rohde, Daniel; Sievers, Hans-Hinrich

    2012-05-01

    An IgG and granulocyte-activating immune response with secondary dystrophic calcification might be the reason glutaraldehyde (GA)-fixed xenograft valves fail, especially in young patients, who are more immunocompetent than the elderly. Titanium nanocoating on GA-fixed bovine pericardium was tested for its ability to prevent major immunoreactions. The immune activity of platelets from GA-fixed bovine pericardium with different treatment procedures was evaluated using the blood from 5 human donors: group I (n = 5), GA fixed as the control; group 2 (n = 5), detoxified with 10% citric acid; group 3 (n = 5), 10% citric acid, aldehyde-dehydrogenase, and a physical plasma treatment; and group 4 (n = 5), treated the same as group 3, but with an additional titanium coat 30 nm in thickness. Titanium deposition was visualized using scanning electron microscopy. IgG deposits (iC3b) were shown by immunostaining and documented as colored pixels (red). The pixels were evaluated electronically. Attracted granulocytes (polymorphonuclear leukocytes) were counted in front of the titanium-coated surface. IC3b deposits and polymorphonuclear leukocytes within control group 1 were defined as 100%; in group 2, iC3b was 149% ± 34% and polymorphonuclear leukocytes were 89%, in group 3, IC3b was 102% ± 24% and polymorphonuclear leukocytes were 47%; and in group 4, IC3b had decreased to 38.49% ± 21% (P < .05) and polymorphonuclear leukocyte activation had decreased to 6.3% (P ≤ .01). Titanium coating significantly reduced the iC3b and granulocyte activating immune response of GA-fixed pericardium. Therefore, it might prevent relevant immunorejection and increase the durability of GA-fixed bioprosthetic heart valves. Copyright © 2012 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.

  13. Effect of penetrating ionising radiation on the mechanical properties of pericardium

    NASA Astrophysics Data System (ADS)

    Daar, Eman; Woods, E.; Keddie, J. L.; Nisbet, A.; Bradley, D. A.

    2010-07-01

    The pericardium is an anistropic composite material made up of collagen and elastin fibres embedded in an amorphous matrix mainly composed of proteoglycan and hyaluronan. The collagen fibres are arranged in layers, with different directions of alignment in each layer, giving rise to interesting mechanical properties of pericardium, including the ability to undergo large deformation during performance of regular physiological functions. The present study aims to investigate the effect of penetrating photon ionising radiation on bovine pericardium tissue, being part of a study of the effect of cardiac doses received in breast radiotherapy and the possibility that this can give rise to cardiovascular complications. Irradiation doses in the range 5-80 Gy were used. To characterise the various mechanical properties [elastic modulus, stress relaxation, ultimate tensile strength (UTS) and fracture] a uniaxial tensile test method was applied. The preliminary results reflect the wide inter-sample variations that are expected in dealing with tissues, with only a weak indication of increase in the UTS of the pericardium tissue with increase in radiation dose. Such an effect has also been observed by others, with reduction in UTS at doses of 80 Gy.

  14. Is Autologous or Heterologous Pericardium Better for Valvuloplasty? A Comparative Study of Calcification Propensity

    PubMed Central

    Jiang, Wen-Jian; Cui, Yong-Chao; Li, Jin-Hua; Zhang, Xiu-Hui; Ding, Huan-Huan; Zhang, Hong-Jia

    2015-01-01

    Pericardial calcification is detrimental to the long-term durability of valvuloplasty. However, whether calcification susceptibility differs between heterologous and autologous pericardium is unclear. In this study, we compared the progression of calcification in vivo between autologous and heterologous pericardium. We randomly divided 28 rabbits into 4 equal groups. Resected rabbit pericardium served as autologous pericardium, and commercial bovine pericardium served as heterologous pericardium. We subcutaneously embedded one of each pericardial patch in the abdominal walls of 21 of the rabbits. The 7 control rabbits (group A) received no implants. The embedded samples were removed at 2 months in group B, at 4 months in group C, and at 6 months in group D. Each collected sample was divided into 2 parts, one for calcium-content measurement by means of atomic-absorption spectroscopy, and one for morphologic and histopathologic examinations. When compared with the autologous pericardium, calcium levels in the heterologous pericardium were higher in groups B, C, and D (P <0.0001, P <0.0002, and P <0.0006, respectively). As embedding time increased, calcium levels in the heterologous pericardium increased faster than those in the autologous, especially in group D. Disorganized arrangements of collagenous fibers, marked calculus, and ossification were seen in the heterologous pericardium. Inflammatory cells—mainly lymphocytes and small numbers of macrophages—infiltrated the heterologous pericardium. The autologous pericardium showed a stronger ability to resist calcification. Our results indicate that autologous pericardium might be a relatively better choice for valvuloplasty. PMID:26175630

  15. Is autologous or heterologous pericardium better for valvuloplasty? A comparative study of calcification propensity.

    PubMed

    Jiang, Wen-Jian; Cui, Yong-Chao; Li, Jin-Hua; Zhang, Xiu-Hui; Ding, Huan-Huan; Lai, Yong-Qiang; Zhang, Hong-Jia

    2015-06-01

    Pericardial calcification is detrimental to the long-term durability of valvuloplasty. However, whether calcification susceptibility differs between heterologous and autologous pericardium is unclear. In this study, we compared the progression of calcification in vivo between autologous and heterologous pericardium. We randomly divided 28 rabbits into 4 equal groups. Resected rabbit pericardium served as autologous pericardium, and commercial bovine pericardium served as heterologous pericardium. We subcutaneously embedded one of each pericardial patch in the abdominal walls of 21 of the rabbits. The 7 control rabbits (group A) received no implants. The embedded samples were removed at 2 months in group B, at 4 months in group C, and at 6 months in group D. Each collected sample was divided into 2 parts, one for calcium-content measurement by means of atomic-absorption spectroscopy, and one for morphologic and histopathologic examinations. When compared with the autologous pericardium, calcium levels in the heterologous pericardium were higher in groups B, C, and D (P <0.0001, P <0.0002, and P <0.0006, respectively). As embedding time increased, calcium levels in the heterologous pericardium increased faster than those in the autologous, especially in group D. Disorganized arrangements of collagenous fibers, marked calculus, and ossification were seen in the heterologous pericardium. Inflammatory cells-mainly lymphocytes and small numbers of macrophages-infiltrated the heterologous pericardium. The autologous pericardium showed a stronger ability to resist calcification. Our results indicate that autologous pericardium might be a relatively better choice for valvuloplasty.

  16. IN VIVO BIOMECHANICAL ASSESSMENT OF TRIGYCIDYLAMINE CROSSLINKED PERICARDIUM

    PubMed Central

    Sacks, Michael S.; Gorman, Robert C.; Hamamoto, Hirotsugu; Connolly, Jeanne M.; Gorman, Joseph H.; Levy, Robert

    2007-01-01

    While a glutaraldehyde crosslinking is most often used to fabricate bioprosthetic heart valves (BHV) using heterograft tissues, it predisposes BHV to calcification and dramatically stiffens the heterograft tissues. Our group previously reported the synthesis and characterization of a novel epoxy-crosslinker, triglycidylamine (TGA). TGA pretreatment of BHV tissues compared to glutaraldehyde results in both calcification resistance in subdermal implants and improved leaflet compliance. In these prior studies, optimal calcification inhibition was noted with the combined use of TGA with mercapto-aminobisphosphonate (MABP). In the present study, we investigated the hypothesis that bovine pericardium cross-linked with TGA-MABP retains these beneficial biomechanical properties in-vivo using a novel mitral valve anterior leaflet (MVAL) ovine valvuloplasty model. Bovine pericardial specimens were crosslinked with either glutaraldehyde or TGA-MABP, from which 1 cm2 sections were implanted in the ovine MVAL after removal of the original tissue of the same size. An array of four sonomicrometry transducers were implanted on the corners and used to compute the complete in-surface strain tensor cardiac cycle over the cardiac cycle at 0 and 4 weeks. Following explant samples were fixed in formalin for histology studies. At 4 weeks both treatment groups experienced no dimensional changes in the unloaded state, indicating no shrinkage. When fully loaded during peak systolic ejection, TGA-MABP valvuloplasty patches were significantly more compliant, which did not change at 4 weeks. In contrast, the glutaraldehyde areal strain increased significantly by 4 weeks. Estimated implant stresses for both treatment groups, based on previously measured biomechanical properties (Biomaterials, vol. 4, 2007), were 40 kPa and 250 kPa in the circumferential and radial directions, respectively, which are comparable to predicted BHV peak stress levels. We conclude that TGA-MABP crosslinked bovine

  17. Donkey pericardium as an alternative bioprosthetic heart valve material.

    PubMed

    Chen, Shanliang; Xu, Li; Liu, Yuxi; Li, Quan; Wang, Dong; Wang, Xuemei; Liu, Tianqi

    2013-03-01

    This study comparatively evaluates the characteristics of glutaraldehyde-treated acellular bovine and donkey pericardium using histological and electronic microscopic observation techniques, shrinkage temperature, and mechanical properties, as well as determining calcium and phosphorus content at 4 and 8 weeks after the subcutaneous implantation of donkey and bovine pericardium in Wistar rats. Donkey pericardium was significantly thinner compared with bovine pericardium (1.622 ±  0.161 mm vs. 4.027 ± 0.401 mm, P < 0.0001) and was associated with significantly greater tensile strength (14.21 ±  3.81 MPa vs. 3.78 ± 1.20 MPa, P = 0.001) and elastic modulus (81.67 ± 20.41 MPa vs. 21.67 ± 11.69 MPa, P <  0.0001) over bovine pericardium. Shrinkage temperature of donkey pericardium was similar to that of bovine pericardium (87.43 ± 0.55°C vs. 87.50 ± 0.36°C, P =  0.810). No differences between groups were observed for maximum load (donkey: 21.64 ± 7.02 KN/m vs. bovine: 15.05 ± 4.50 KN/m, P = 0.082) and tear strength (donkey: 11.54 ± 5.33 MPa vs. bovine: 10.69 ±  3.77 MPa, P = 0.757). Calcium content was significantly lower in donkey pericardium compared with bovine pericardium at 4 weeks (690.15 ± 191.27 µg/g vs. 1381.73 ± 62.52 µg/g, P = 0.001) and 8 weeks (205.24 ± 62.40 µg/g vs. 910.48 ± 398.29 µg/g, P = 0.037). This preliminary study has confirmed that glutaraldehyde-tanned donkey pericardium, demonstrating reduced calcification and increased tensile strength, may provide a suitable bioprosthetic valve substitute.

  18. Dynamic compressive properties of bovine knee layered tissue

    NASA Astrophysics Data System (ADS)

    Nishida, Masahiro; Hino, Yuki; Todo, Mitsugu

    2015-09-01

    In Japan, the most common articular disease is knee osteoarthritis. Among many treatment methodologies, tissue engineering and regenerative medicine have recently received a lot of attention. In this field, cells and scaffolds are important, both ex vivo and in vivo. From the viewpoint of effective treatment, in addition to histological features, the compatibility of mechanical properties is also important. In this study, the dynamic and static compressive properties of bovine articular cartilage-cancellous bone layered tissue were measured using a universal testing machine and a split Hopkinson pressure bar method. The compressive behaviors of bovine articular cartilage-cancellous bone layered tissue were examined. The effects of strain rate on the maximum stress and the slope of stress-strain curves of the bovine articular cartilage-cancellous bone layered tissue were discussed.

  19. Differential expression of Dlk-1 in bovine adipose tissue depots.

    PubMed

    Vuocolo, T; Pearson, R; Campbell, P; Tellam, R L

    2003-02-01

    Dlk-1, a type 1 membrane glycoprotein, is a member of the Epidermal Growth Factor-like family of homeotic proteins that are typically involved in cell fate decisions and in mice it has been implicated in the control of differentiation of adipocytes. The aim of this study was to determine whether there were tissue-specific expression patterns of Dlk-1 splice variants in bovine tissues. Only the Dlk-1-C2 variant was expressed in adult bovine tissues while both Dlk-1-C2 and Dlk-1-A variants were expressed in foetal tissues. Quantitative real-time PCR revealed large differences in the relative levels of expression of the Dlk-1-C2 variant in adult adipose tissue depots with no expression in subcutaneous and brisket adipose tissues. Expression was also demonstrated in three adult skeletal muscle samples. The large variation in the level of expression of Dlk-1-C2 in different adult tissues may reflect the relative preadipocyte content of those tissues and consequently their potential for generating new adipocytes. A low abundance soluble glycoprotein (bFA1) was purified from bovine amniotic fluid. Analyses of its amino acid sequence revealed that it corresponded to most of the extracellular domain of bovine Dlk-1 and was derived by proteolytic processing from the full-length Dlk-1 protein encoded by the Dlk-1-A variant. The tissues expressing the Dlk-1-A variant have not been identified but are likely to be foetal in origin. Splice variants of Dlk-1 may have varied functional roles with the foetal Dlk-1-A form capable of generating a protein that undergoes proteolytic processing to release a soluble ecto-domain of Dlk-1. In contrast the Dlk-1-C2 splice variant codes for a protein lacking this processing site and therefore it probably remains bound to the cell membrane.

  20. Bovine Leukemia Virus DNA in Human Breast Tissue

    PubMed Central

    Shen, Hua Min; Jensen, Hanne M.; Choi, K. Yeon; Sun, Dejun; Nuovo, Gerard

    2014-01-01

    Bovine leukemia virus (BLV), a deltaretrovirus, causes B-cell leukemia/lymphoma in cattle and is prevalent in herds globally. A previous finding of antibodies against BLV in humans led us to examine the possibility of human infection with BLV. We focused on breast tissue because, in cattle, BLV DNA and protein have been found to be more abundant in mammary epithelium than in lymphocytes. In human breast tissue specimens, we identified BLV DNA by using nested liquid-phase PCR and DNA sequencing. Variations from the bovine reference sequence were infrequent and limited to base substitutions. In situ PCR and immunohistochemical testing localized BLV to the secretory epithelium of the breast. Our finding of BLV in human tissues indicates a risk for the acquisition and proliferation of this virus in humans. Further research is needed to determine whether BLV may play a direct role in human disease. PMID:24750974

  1. Bovine leukemia virus DNA in human breast tissue.

    PubMed

    Buehring, Gertrude Case; Shen, Hua Min; Jensen, Hanne M; Choi, K Yeon; Sun, Dejun; Nuovo, Gerard

    2014-05-01

    Bovine leukemia virus (BLV), a deltaretrovirus, causes B-cell leukemia/lymphoma in cattle and is prevalent in herds globally. A previous finding of antibodies against BLV in humans led us to examine the possibility of human infection with BLV. We focused on breast tissue because, in cattle, BLV DNA and protein have been found to be more abundant in mammary epithelium than in lymphocytes. In human breast tissue specimens, we identified BLV DNA by using nested liquid-phase PCR and DNA sequencing. Variations from the bovine reference sequence were infrequent and limited to base substitutions. In situ PCR and immunohistochemical testing localized BLV to the secretory epithelium of the breast. Our finding of BLV in human tissues indicates a risk for the acquisition and proliferation of this virus in humans. Further research is needed to determine whether BLV may play a direct role in human disease.

  2. A cellular mechanism for imidocarb retention in edible bovine tissues.

    PubMed

    Moore, A S; Coldham, N G; Sauer, M J

    1996-10-01

    Imidocarb dipropionate, formulated as Imizol, is used for the treatment and prophylaxis of bovine babesiosis. Several studies have shown that imidocarb remains detectable in edible ovine and bovine tissues for several months after dosing but the mechanism of retention remains unknown. In this study, the mechanism of imidocarb retention was investigated by measuring the binding of [14C]imidocarb to bovine hepatocytes, erythrocytes, sub-cellular fractions and isolated bovine macromolecules. The proportion of [14C]imidocarb (10 microM) bound to cells in suspension culture (1 x 10(7) cells.ml-1) was found to be substantially greater to hepatocytes (56.5%) than to erythrocytes (4.6%). Studies with washed erythrocytes reconstituted in plasma indicated that approximately 70% of the [14C]imidocarb was bound to plasma proteins, 10% to erythrocytes, and 20% remained free. Measurement of [14C]imidocarb binding to sub-cellular fractions prepared from bovine liver revealed preferential accumulation in the nuclear, rather than in the mitochondrial, microsomal or cytosolic fractions. Binding capacities of selected bovine macromolecules for [14C]imidocarb were in the order deoxy-ribonucleic acid (DNA) = ribonucleic acid (RNA) > > alpha 1-acid glycoprotein (AGP) > serum albumin (BSA) > haemoglobin (Hb). DNA binding sites for imidocarb remained unsaturated over the concentration range 0-100 microM [14C]imidocarb. Competitive binding studies between imidocarb and pentamidine or spermidine provided evidence for common DNA binding sites. These studies indicated that preferential binding of [14C]imidocarb to hepatocytes compared with erythrocytes observed in vitro was a result of substantial reversible binding to nucleic acids and that the same cellular mechanism may be implicated in the slow elimination of imidocarb from edible tissues in vivo.

  3. Expression of a 50 kDa putative receptor for bovine viral diarrhea virus in bovine fetal tissues.

    PubMed Central

    Zheng, L; Zhang, S; Xue, W; Kapil, S; Minocha, H C

    1998-01-01

    The expression of a 50 kDa bovine viral diarrhea virus putative receptor in different bovine fetal tissues from 3-month old fetuses was studied. The receptor expression was examined by immunocytochemical staining and by immunoblotting using antiidiotypic probe (anti-D89). Intense specific staining in enterocytes of the small and large intestines, cortical tubular epithelial cells of kidneys, respiratory epithelial cells of the trachea and esophageal mucosal epithelial cells was observed, demonstrating the strong expression of bovine viral diarrhea virus receptor in the tissues. Weak staining was found in cerebellum, thymus, spleen, liver, cerebrum, and lung tissues; however, heart tissues were negative. Immunoblotting results correlated with the immunoperoxidase staining assays. Thus, the expression levels of the receptor are variable in different tissues. This pattern of expression may provide clues to the pathogenic potential of bovine viral diarrhea virus in the bovine fetus. Images Figure 1. Figure 2. PMID:9553718

  4. The effects of the covalent attachment of 3-(4-hydroxy-3,5-di-tert-butylphenyl)propyl amine to glutaraldehyde pre-treated bovine pericardium on structural degeneration, oxidative modification and calcification of rat subdermal implants

    PubMed Central

    Christian, Abigail J.; Alferiev, Ivan S.; Connolly, Jeanne M.; Ischiropoulos, Harry; Levy, Robert J.

    2014-01-01

    Bioprosthetic heart valves (BHV) fabricated from glutaraldehyde pretreated heterograft materials, porcine aortic valves or bovine pericardium (BP), are widely used in cardiac surgery. BHV progressively fail in clinical use due to structural degeneration. Previously we reported that dityrosine, an oxidized amino acid, was present in failed clinical BP-BHV explants; unimplanted BP had no detectable dityrosine. In the same studies BP were demonstrated in vitro to be susceptible to oxidative damage, that could be mitigated with BP covalently modified with the antioxidant, 3-(4-hydroxy-3,5-di-tert-butylphenyl)propyl amine (DBP). The present studies compared in rat subdermal implants glutaraldehyde pretreated BP to BP modified with either DBP or the chemical reactions used to link DBP. All BP explants regardless of DBP demonstrated reduced hydroxyproline and increased digestibility by collagenase. However, the DBP-BP explants showed significant inhibition of reduced explant shrink temperatures (an index of crosslinking) compared to control BP. Significant mitigation of calcification was observed in both the BP-DBP and chemically modified explants compared to BP. Dityrosine was not detectable in the 90 day explants. It is concluded that rat subdermal BP implants undergo both calcific and non-calcific structural degeneration, but without the formation of dityrosine, unlike clinical BP explants. PMID:25546235

  5. The effects of the covalent attachment of 3-(4-hydroxy-3,5-di-tert-butylphenyl) propyl amine to glutaraldehyde pretreated bovine pericardium on structural degeneration, oxidative modification, and calcification of rat subdermal implants.

    PubMed

    Christian, Abigail J; Alferiev, Ivan S; Connolly, Jeanne M; Ischiropoulos, Harry; Levy, Robert J

    2015-07-01

    Bioprosthetic heart valves (BHV) fabricated from glutaraldehyde pretreated heterograft materials, porcine aortic valves or bovine pericardium (BP), are widely used in cardiac surgery. BHV progressively fail in clinical use due to structural degeneration. Previously we reported that dityrosine, an oxidized amino acid, was present in failed clinical BP-BHV explants; unimplanted BP had no detectable dityrosine. In the same studies BP were demonstrated in vitro to be susceptible to oxidative damage, that could be mitigated with BP covalently modified with the antioxidant, 3-(4-hydroxy-3,5-di-tert-butylphenyl)propyl amine (DBP). The present studies compared in rat subdermal implants glutaraldehyde pretreated BP to BP modified with either DBP or the chemical reactions used to link DBP. All BP explants regardless of DBP demonstrated reduced hydroxyproline and increased digestibility by collagenase. However, the DBP-BP explants showed significant inhibition of reduced explant shrink temperatures (an index of crosslinking) as compared with control BP. Significant mitigation of calcification was observed in both the BP-DBP and chemically modified explants as compared with BP. Dityrosine was not detectable in the 90 day explants. It is concluded that rat subdermal BP implants undergo both calcific and noncalcific structural degeneration, but without the formation of dityrosine, unlike clinical BP explants. © 2014 Wiley Periodicals, Inc.

  6. Native and recombinant bovine growth hormone antagonize insulin action in cultured bovine adipose tissue.

    PubMed

    Etherton, T D; Evock, C M; Kensinger, R S

    1987-08-01

    The current study was undertaken to determine if pituitary bovine GH (pbGH) and recombinant bGH (rbGH) antagonized insulin action in bovine adipose tissue after acute (2-h) and chronic (48-h) exposure and whether this was an intrinsic property of bGH. Insulin action (measured as the effect on incorporation of acetate-carbon into long-chain fatty acids) was unaffected by bGH in short term incubations regardless of whether hydrocortisone (HC) was present. After 48 h of culture, however, both pbGH and rbGH similarly antagonized the ability of insulin to maintain lipogenic capacity. This antagonism was dependent upon the presence of HC and was dose dependent, with half-maximal inhibition of insulin action occurring at about 0.5 ng/ml bGH. Bovine PRL did not mimic the effects of bGH on insulin action. These results establish that bGH antagonizes insulin action in bovine adipose tissue and that this effect is dependent upon long term exposure and the inclusion of HC in the culture medium. The fact that both rbGH and pbGH acted similarly indicates that this is an intrinsic property of bGH. The effect of bGH on insulin-dependent maintenance of lipogenic capacity may play an important role in redirecting nutrients away from adipose tissue to other tissues, such as muscle or mammary tissue. It is speculated that this metabolic effect of bGH plays an important role in the adaptive response to chronic bGH treatment, which increases milk yield of dairy cows and growth performance of beef cattle.

  7. Alcohol-based solutions for bovine testicular tissue fixation.

    PubMed

    Cabrera, Nelson C; Espinoza, Jorge R; Vargas-Jentzsch, Paul; Sandoval, Patricio; Ramos, Luis A; Aponte, Pedro M

    2017-01-01

    Tissue fixation, a central element in histotechnology, is currently performed with chemical compounds potentially harmful for human health and the environment. Therefore, alternative fixatives are being developed, including alcohol-based solutions. We evaluated several ethanol-based mixtures with additives to study fixative penetration rate, tissue volume changes, and morphologic effects in the bovine testis. Fixatives used were Bouin solution, 4% formaldehyde (F4), 70% ethanol (E70), E70 with 1.5% glycerol (E70G), E70 with 5% acetic acid (E70A), E70 with 1.5% glycerol and 5% acetic acid (E70AG), and E70 with 1.5% glycerol, 5% acetic acid, and 1% dimethyl sulfoxide (DMSO; E70AGD). Five-millimeter bovine testicular tissue cubes could be completely penetrated by ethanol-based fixatives and Bouin solution in 2-3 h, whereas F4 required 21 h. Bouin solution produced general tissue shrinkage, whereas the other fixatives (alcohol-based and F4) caused tissue volume expansion. Although Bouin solution is an excellent fixative for testicular tissue, ethanol-based fixatives showed good penetration rates, low tissue shrinkage, and preserved sufficient morphology to allow identification of the stages of the seminiferous epithelium cycle, therefore representing a valid alternative for histotechnology laboratories. Common additives such as acetic acid, glycerol, and DMSO offered marginal benefits for the process of fixation; E70AG showed the best preservation of morphology with excellent nuclear detail, close to that of Bouin solution.

  8. Pathophysiology of the Pericardium.

    PubMed

    Hoit, Brian D

    Pericardial heart disease includes pericarditis, (an acute, subacute, or chronic fibrinous, noneffusive, or exudative process), and its complications, constriction, (an acute, subacute, or chronic adhesive or fibrocalcific response), and cardiac tamponade. The pathophysiology of cardiac tamponade and constrictive pericarditis readily explains their respective findings on clinical examination, Doppler echocardiography, and at cardiac catheterization. The primary abnormality of cardiac tamponade is pan-cyclic compression of the cardiac chambers by increased pericardial fluid requiring that cardiac chambers compete for a fixed intrapericardial volume. Features responsible for the pathophysiology include transmission of thoracic pressure through the pericardium and heightened ventricular interdependence. Constrictive pericarditis is a condition in which the pericardium limits diastolic filling and causes dissociation of intracardiac and intrathoracic pressures, and heightened ventricular interdependence. Both conditions result in diastolic dysfunction, elevated and equal venous and ventricular diastolic pressure, respiratory variation in ventricular filling, and ultimately, reduced cardiac output. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Biostability, durability and calcification of cryopreserved human pericardium after rapid glutaraldehyde-stabilization versus multistep ADAPT(R) treatment in a subcutaneous rat model.

    PubMed

    Neethling, William; Brizard, Christian; Firth, Laura; Glancy, Ross

    2014-04-01

    Autologous pericardium rapidly fixed with glutaraldehyde (GA) in theatre is considered in many cardiac surgery centres the best material currently available for intracardiac, valvular or vascular repair. Implanted non-fixed autologous tissues suffer rapid degeneration, shrinkage and absorption whereas standard xenotypic fixed tissues cause local cytotoxicity and calcification. In the present study, using a subcutaneous rat model, we tested the biostability, durability and calcification potential of four different pericardium patches treated with GA and relevant to current clinical practice. Pericardium samples were divided into four groups according to the method of treatment. Group I consisted of bovine pericardium (BP) fixed with 0.6% GA (control), Group II cryopreserved human pericardium (CHP) rapidly fixed with 0.6% GA for 4 min and detoxified with MgCl2, Group III CHP treated with the multistep ADAPT(®) process (delipidized, decellularized with Tx-100, deoxycholate, IgePal CA-630 and denucleased, fixed in 0.05% monomeric GA and detoxified) and Group IV BP treated with the multistep ADAPT(®) process (CardioCel(®)). Biostability was determined by shrinkage temperature which measures the degree of cross-linking, and durability assessed by resistance to a mixture of proteinases (pronase digestion). Treated pericardium samples (n = 10 in each of Groups I-IV) were implanted in the subcutaneous rat model for 8 and 16 weeks, followed by histology and calcium analysis (atomic absorption spectrophotometry). The biostability and the durability of both CHP and BP after the multistep ADAPT(®) treatment remained stable without any microscopic calcification. Extractable calcium levels of CHP were significantly (P < 0.01) reduced in Group II (1.89 ± 0.77 μg Ca/mg tissue) compared with Group I (64.37 ± 6.25 μg/mg) after 8 weeks. Calcification of CHP (Group III) and BP (Group IV) after the multistep ADAPT(®) treatment was significantly reduced (1.43 ± 0.48 µg/mg and

  10. Congenital Absence of the Pericardium.

    PubMed

    Lopez, David; Asher, Craig R

    Congenital absence of the pericardium (CAP) is one of the rarest cardiac congenital anomalies. It can occur as a complete absence of the entire pericardium, absence of the right or left portion of the pericardium or a partial, foramen-like defect of the right or left pericardium. While the majority of cases are clinically silent, multiple reports associate CAP with symptomatic presentation. The most feared complication of CAP is sudden death due to cardiac strangulation across a partial defect of the left pericardium. Given its rare occurrence, most clinicians and imaging specialists will have little experience with this condition and may fail to recognize it on thoracic or cardiac studies. Thus, the purpose of this review is to highlight the common clinical and multimodality imaging features associated with this anomaly and suggest a management algorithm. Copyright © 2016. Published by Elsevier Inc.

  11. In vitro metabolism of ceftiofur in bovine tissues.

    PubMed

    Olson, S C; Beconi-Barker, M G; Smith, E B; Martin, R A; Vidmar, T J; Adams, L D

    1998-04-01

    The metabolism of ceftiofur in bovine kidney, liver, muscle and lung, and the effects of the presence of cystine and glutathione in the media were evaluated using S-9 and microsomal tissue fractions. Conversion of ceftiofur to desfuroylceftiofur (DFC) was catalyzed by an esterase which was most active in kidney, followed by liver. It was not very active in muscle and lung. After DFC was liberated, it rapidly bound primarily to tissue proteins (> 56%), and was also conjugated to cysteine and glutathione. Production of DFC-cysteine by disulfide exchange of DFC with cystine and production of DFC-glutathione by conjugation of DFC to glutathione occurred in buffer if glutathione and cystine were present in the medium. These conjugations were also observed in incubations with tissue fractions, indicating that they were not inhibited by the tissues endogenous molecules. In addition, the metabolism of DFC-glutathione to DFC-cysteine was observed when tissue proteins were present. The metabolism of DFC-glutathione to DFC-cysteine was faster in kidney than in liver. Metabolites devoid of an intact beta-lactam ring were not observed in these in vitro studies.

  12. Pericardium: structure and function in health and disease.

    PubMed

    Jaworska-Wilczynska, Maria; Trzaskoma, Pawel; Szczepankiewicz, Andrzej A; Hryniewiecki, Tomasz

    2016-01-01

    Normal pericardium consists of an outer sac called fibrous pericardium and an inner one called serous pericardium. The two layers of serous pericardium: visceral and parietal are separated by the pericardial cavity, which contains 20 to 60 mL of the plasma ultrafiltrate. The pericardium acts as mechanical protection for the heart and big vessels, and a lubrication to reduce friction between the heart and the surrounding structures. A very important role in all aspects of pericardial functions is played by mesothelial cells. The mesothelial cells form a monolayer lining the serosal cavity and play an important role in antigen presentation, inflammation and tissue repair, coagulation and fibrinolysis. The two major types of mesothelial cells, flat or cuboid, differ substantially in their ultrastructure and, probably, functions. The latter display abundant microvilli, RER, Golgi dense bodies, membrane-bound vesicles and intracellular vacuoles containing electron-dense material described as dense bodies. The normal structure and functions of the pericardium determine correct healing after its injury as a result of surgery or microbial infection. The unfavorable resolution of acute or chronic pericarditis leads to the formation of adhesions between pericardial leaflets which may lead to serious complications.

  13. Usefulness of the transgastric view by transesophageal echocardiography in evaluating thickened pericardium in patients with constrictive pericarditis.

    PubMed

    Izumi, Chisato; Iga, Kanji; Sekiguchi, Kiyomi; Takahashi, Shuichi; Konishi, Takashi

    2002-09-01

    Detection of thickened pericardium in patients with constrictive pericarditis is essential for pericardiectomy because restrictive cardiomyopathy and severe tricuspid regurgitation show similar hemodynamic data. The purpose of this study was to clarify whether transesophageal echocardiography can evaluate thickened pericardium. We investigated 7 patients with constrictive pericarditis who underwent pericardiectomy. Thickened pericardium over the right atrium was detected in 6 patients, but the borders were not clear. Thickened pericardium over the left ventricle was not detected in any patients in the standard longitudinal and horizontal views. On the other hand, thickened pericardium over the ventricles was detected in all patients in the transgastric view as an echogenic area between the liver and ventricular wall. Tissue characteristics of the thickened pericardium could be evaluated because of the high-quality images in the transgastric view. The transgastric view by transesophageal echocardiography allows high-quality images of the pericardium, which might be useful in diagnosing constrictive pericarditis.

  14. [Dehiscence of the Pericardial Patch, after Surgical Treatment of Active Infective Aortic Valve Endocarditis with Reconstruction of the Aortic Annulus Using a Glutaraldehyde-treated Autologous Pericardium].

    PubMed

    Yoshida, Kazufumi; Fukunaga, Naoto; Koizumi, Shigeki; Nishiya, Kenta; Matsuda, Yasuhiro; Ishigami, Masanosuke; Nagasawa, Atsushi; Sakata, Ryuzo; Koyama, Tadaaki

    2017-03-01

    A 50-year-old man was admitted with fever and chill sensation 6 months ago. Transthoracic echocardiography (TTE) showed left and right coronary cusp prolapses and a thickened tissue of the aortic curtain. Congestive heart failure due to active infective aortic valve endocarditis was diagnosed, and he underwent aortic valve replacement. The aortic annulus was reconstructed using a glutaraldehydetreated autologous pericardium. Six months after surgery, TTE showed severe aortic regurgitation and saccular change in the aortic annulus. Transesophageal echocargiography showed some echo free space from left to noncoronary cusp and abnormal movement of the prosthetic valve annulus. Intraoperative examination showed dehiscence of the pericardial patch from the aortic wall, but no finding of infection. Aortic valve rereplacement was performed with reconstruction of the aortic annulus using a bovine pericardium. To prevent the dehiscence of the pericardial patch from the aortic wall, sutures fixing the prosthetic valve were passed from outside of the aortic wall.

  15. Follicle Viability after Vitrification of Bovine Ovarian Tissue.

    PubMed

    Guedes, Janaína de Souza; Rodrigues, Jhenifer Kliemchen; Campos, Ana Luisa Menezes; Moraes, Camila Cruz de; Caetano, João Pedro Junqueira; Marinho, Ricardo Mello

    2017-08-31

    Purpose The present study aimed to evaluate the impact of vitrification on the viability of follicles using a three-dimensional (3D) in vitro culture. Methods Bovine ovarian tissue samples (n = 5) obtained from slaughterhouses were utilized. The cortex was cut into small fragments of 2 × 3 × 0.5 mm using a tissue slicer. From these fragments, secondary follicles were first isolated by mechanical and enzymatic methods, then encapsulated in alginate gel and individually cultured for 20 days. Additional fragments of the same ovarian tissue were vitrified in a solution containing 25% glycerol and 25% ethylene glycol. After warming, the follicles underwent the same follicular isolation process that was performed for the fresh follicles. Results A total of 61 follicles were isolated, 51 from fresh ovarian tissue, and 10 from vitrified tissue. After the culture, the vitrified and fresh follicles showed 20% and 43.1% survival rates respectively (p = 0.290), with no significant differences. At the end of the culture, there were no significant differences in follicular diameter between the vitrified (422.93 ± 85.05 µm) and fresh (412.99 ± 102.55 µm) groups (p = 0.725). Fresh follicles showed higher mean rate of antrum formation when compared with vitrified follicles (47.1% and 20.0% respectively), but without significant difference (p = 0.167). Conclusions The follicles were able to develop, grow and form antrum in the 3D system after vitrification, despite the lower results obtained with the fresh tissue. Thieme Revinter Publicações Ltda Rio de Janeiro, Brazil.

  16. Enzymes involved in acetoacetate formation in various bovine tissues.

    PubMed

    Baird, G D; Hibbitt, K G; Lee, J

    1970-05-01

    1. The activities of acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA synthase and lyase and acetoacetyl-CoA deacylase were measured in homogenates of samples of liver, rumen epithelium (long papillae), kidney and lactating mammary gland derived from slaughtered cows. 2. The activities of the four enzymes in bovine liver were similar to the activities previously reported for the corresponding enzymes in rat liver. 3. Acetoacetyl-CoA thiolase and hydroxymethylglutaryl-CoA synthase and lyase were present in rumen epithelium. The activities of the enzymes were all lower on a wet weight basis than in liver. Only very slight deacylase activity was detected. 4. Kidney contained acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA lyase and acetoacetyl-CoA deacylase, but only trace amounts of hydroxymethylglutaryl-CoA synthase. 5. Mammary gland contained acetoacetyl-CoA thiolase and some hydroxymethylglutaryl-CoA lyase, but virtually no hydroxymethylglutaryl-CoA synthase or acetoacetyl-CoA deacylase. 6. Since physiologically significant ketogenesis probably occurs solely via the hydroxymethylglutaryl-CoA pathway, it is evident that, of the four tissues examined, such ketogenesis must be restricted to the liver and the rumen epithelium. 7. All the enzymes except hydroxymethylglutaryl-CoA lyase were also assayed in the four tissues derived from cows suffering from bovine lactational ketosis. Ketosis did not cause a statistically significant change in the activity of any of the enzymes measured. 8. Hepatic hydroxymethylglutaryl-CoA synthase and lyase were found to be associated mainly with the particulate fraction, as in the rat. A considerably greater proportion of these enzymes was found to be present in the cytoplasmic fraction from rumen epithelium, although it was not excluded that this was due to mitochondrial damage during homogenization. 9. Appreciable hydroxymethylglutaryl-CoA synthase was also present in epithelium from the dorsal region of the rumen, from the

  17. Simulation of the contraction of the ventricles in a human heart model including atria and pericardium.

    PubMed

    Fritz, Thomas; Wieners, Christian; Seemann, Gunnar; Steen, Henning; Dössel, Olaf

    2014-06-01

    During the contraction of the ventricles, the ventricles interact with the atria as well as with the pericardium and the surrounding tissue in which the heart is embedded. The atria are stretched, and the atrioventricular plane moves toward the apex. The atrioventricular plane displacement (AVPD) is considered to be a major contributor to the ventricular function, and a reduced AVPD is strongly related to heart failure. At the same time, the epicardium slides almost frictionlessly on the pericardium with permanent contact. Although the interaction between the ventricles, the atria and the pericardium plays an important role for the deformation of the heart, this aspect is usually not considered in computational models. In this work, we present an electromechanical model of the heart, which takes into account the interaction between ventricles, pericardium and atria and allows to reproduce the AVPD. To solve the contact problem of epicardium and pericardium, a contact handling algorithm based on penalty formulation was developed, which ensures frictionless and permanent contact. Two simulations of the ventricular contraction were conducted, one with contact handling of pericardium and heart and one without. In the simulation with contact handling, the atria were stretched during the contraction of the ventricles, while, due to the permanent contact with the pericardium, their volume increased. In contrast to that, in the simulations without pericardium, the atria were also stretched, but the change in the atrial volume was much smaller. Furthermore, the pericardium reduced the radial contraction of the ventricles and at the same time increased the AVPD.

  18. The role of the pericardium in the valveless, tubular heart of the tunicate Ciona savignyi.

    PubMed

    Waldrop, Lindsay D; Miller, Laura A

    2015-09-01

    Tunicates, small invertebrates within the phylum Chordata, possess a robust tubular heart which pumps blood through their open circulatory systems without the use of valves. This heart consists of two major components: the tubular myocardium, a flexible layer of myocardial cells that actively contracts to drive fluid down the length of the tube; and the pericardium, a stiff, outer layer of cells that surrounds the myocardium and creates a fluid-filled space between the myocardium and the pericardium. We investigated the role of the pericardium through in vivo manipulations on tunicate hearts and computational simulations of the myocardium and pericardium using the immersed boundary method. Experimental manipulations reveal that damage to the pericardium results in aneurysm-like bulging of the myocardium and major reductions in the net blood flow and percentage closure of the heart's lumen during contraction. In addition, varying the pericardium-to-myocardium (PM) diameter ratio by increasing damage severity was positively correlated with peak dye flow in the heart. Computational simulations mirror the results of varying the PM ratio experimentally. Reducing the stiffness of the myocardium in the simulations reduced mean blood flow only for simulations without a pericardium. These results indicate that the pericardium has the ability to functionally increase the stiffness of the myocardium and limit myocardial aneurysms. The pericardium's function is likely to enhance flow through the highly resistive circulatory system by acting as a support structure in the absence of connective tissue within the myocardium. © 2015. Published by The Company of Biologists Ltd.

  19. Fluorescence of tissues fluororophores such as lipofuscin as a possible basis for the detection of CNS tissues on bovine carcasses

    USDA-ARS?s Scientific Manuscript database

    Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative transmissible spongiform encephalopathy (TSE) which is thought to cause variant of Creutzfeldt-Jakob disease (vCJD) in humans. It is believed that humans contract vCJD by consumption of meat contaminated with bovine tissue containi...

  20. Structure and Anatomy of the Human Pericardium.

    PubMed

    Rodriguez, E Rene; Tan, Carmela D

    The normal gross anatomy and light microscopy of the human pericardium are presented in detail that allows easy correlation with current cardiac imaging modalities. The anatomical structures of the parietal pericardium are shown from its mediastinal surface, including its ligaments to the sternum, diaphragm and vertebral column. The attachments of the parietal pericardium to the great vessels showing the intrapericardial location of the root of the aorta and pulmonary artery are documented. Also the attachments of the parietal pericardium to the venae cavae and the pulmonary veins are illustrated in detail. The internal anatomy of the parietal pericardium emphasizing the oblique and transverse sinuses is explained. The microscopic differences between the structures of the parietal pericardium and visceral pericardium (epicardium) are shown as the basis that allows understanding the spectrum of adaptation of the pericardium to diverse pathologic processes. However, the pathology of the pericardium is not discussed in this review. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. In vitro comparative assessment of decellularized bovine pericardial patches and commercial bioprosthetic heart valves.

    PubMed

    Aguiari, Paola; Iop, Laura; Favaretto, Francesca; Fidalgo, Cátia Marisa Lourenco; Naso, Filippo; Milan, Gabriella; Vindigni, Vincenzo; Spina, Michel; Bassetto, Franco; Bagno, Andrea; Vettor, Roberto; Gerosa, Gino

    2017-02-03

    Notwithstanding their wide exploitation, biological prosthetic heart valves are characterized by limited durability (10-15 years). The treatment of biological tissues with chemical crosslinking agents such as glutaraldehyde accounts for the enhanced risk of structural deterioration associated with the early failure of bioprosthetic valves. To overcome the shortcomings of the currently available solutions, adoption of decellularized biological tissues of animal origin has emerged as a promising approach. The present study aims to assess in vitro cardiovascular scaffolds composed of bovine pericardium decellularized with the novel TRITDOC (TRIton-X100 and TauroDeOxyCholic acid) procedure. The effects of the treatment have been assessed by means of histological, biomolecular, cellular, biochemical and biomechanical analyses. The TRITDOC procedure grants the complete decellularization of bovine pericardial scaffolds while preserving the extracellular matrix architecture and the biomechanical properties. With a dedicated ELISA test, the TRITDOC procedure has been proven to ensure the complete removal of the alphaGal antigen, responsible for hyperacute rejection and for long-term deterioration of xenogenic biomaterials. Static seeding of the acellular pericardial patches with human adipose-derived stem cells resulted in an evenly repopulated scaffold without signs of calcification. The in vitro cyto-/immuno-compatibility response of the TRITDOC-bovine pericardium was compared with glutaraldehyde-treated xenogenic pericardium collected from two bioprosthetic devices currently used in clinical practice: PERIMOUNT MAGNA and TRIFECTA(TM). TRITDOC-bovine pericardium exhibited lower complement activation, lower cytotoxicity and a lower tendency to secrete pro-inflammatory cytokines compared to the tested commercial bioprostheses. Therefore, TRITDOC-decellularized pericardium could be considered as possible candidate material for the production of prosthetic heart valves.

  2. Guidelines for sizing pericardium for aortic valve leaflet grafts

    PubMed Central

    Hammer, Peter E.; del Nido, Pedro J.

    2013-01-01

    Surgical repair of the aortic valve using leaflet grafts made from pericardium has been shown to be a viable option, particularly in children, in whom valve replacement has strong disadvantages. We present guidelines for sizing treated autologous pericardium to fabricate a leaflet graft for single leaflet replacement. Both our clinical experience and experimental evidence indicate that effective repairs are best achieved using a semicircular graft with diameter 10–15% greater than the sinotubular junction diameter in diastole. We also provide a simple formula to allow adjusting these guidelines to account for variation in valve geometry and tissue properties. PMID:23816114

  3. Flexor Tendon Sheath Engineering Using Decellularized Porcine Pericardium.

    PubMed

    Megerle, Kai; Woon, Colin; Kraus, Armin; Raghavan, Shyam; Pham, Hung; Chang, James

    2016-10-01

    The flexor tendon sheath is an ideal target for tissue engineering because it is difficult to reconstruct by conventional surgical methods. The authors hypothesized that decellularized porcine pericardium can be used as a scaffold for engineering a biologically active tendon sheath. The authors' protocol removed cellular material from the pericardium and preserved the structural architecture in addition to the collagen and glycosaminoglycan content. The scaffold was successfully reseeded with human sheath synoviocytes and human adipose-derived stem cells. Cells were evaluated for 8 weeks after reseeding. The reseeded construct demonstrated continuous production of hyaluronic acid, the main component of synovial fluid. After being seeded on the membrane, adipose-derived stem cells demonstrated down-regulation of collagen I and III and up-regulation of hyaluronan synthase 2. The results indicate that decellularized porcine pericardium may be a potential scaffold for engineering a biologically active human tendon sheath.

  4. A dynamic model to calculate cadmium concentrations in bovine tissues from basic soil characteristics.

    PubMed

    Waegeneers, Nadia; Ruttens, Ann; De Temmerman, Ludwig

    2011-06-15

    A chain model was developed to calculate the flow of cadmium from soil, drinking water and feed towards bovine tissues. The data used for model development were tissue Cd concentrations of 57 bovines and Cd concentrations in soil, feed and drinking water, sampled at the farms were the bovines were reared. Validation of the model occurred with a second set of measured tissue Cd concentrations of 93 bovines of which age and farm location were known. The exposure part of the chain model consists of two parts: (1) a soil-plant transfer model, deriving cadmium concentrations in feed from basic soil characteristics (pH and organic matter content) and soil Cd concentrations, and (2) bovine intake calculations, based on typical feed and water consumption patterns for cattle and Cd concentrations in feed and drinking water. The output of the exposure model is an animal-specific average daily Cd intake, which is then taken forward to a kinetic uptake model in which time-dependent Cd concentrations in bovine tissues are calculated. The chain model was able to account for 65%, 42% and 32% of the variation in observed kidney, liver and meat Cd concentrations in the validation study.

  5. Congenital Absence of the Pericardium

    PubMed Central

    Kim, Hyun-Jin; Cho, Goo-Yeong; Choi, Sang Il

    2014-01-01

    Congenital absence of the pericardium is a rare cardiac malformation and is most often asymptomatic. It is usually discovered as an incidental finding. Physical examination, chest radiography, and electrocardiogram are often unremarkable. Echocardiography provides valuable information, and sometimes computed tomography or magnetic resonance imaging is needed for subsequent confirmation. PMID:24753808

  6. Identification of a cDNA clone for bovine tissue factor

    SciTech Connect

    Kittler, J.M.; Horton, R.D.; Bach, R.; Spicer, E.K.; Fitzgerald, M.J.; Nemerson, Y.; Konigsberg, W.H.

    1986-05-01

    Tissue factor is a membrane-bound glycoprotein of approx.43 Kd that is necessary for activation of the extrinsic pathway of blood coagulation. The amino terminal amino acid sequence of purified bovine tissues factor was used to design a synthetic 17-mer oligodeoxyribonucleotide probe. A lambda gtll bovine brain cortex cDNA expression library was screened using both the /sup 32/P-labeled oligodeoxynucleotide probe and polyclonal antibodies to bovine tissue factor. A recombinant phage was isolated which gave a positive reaction with both probes. Cells containing the lambda gtll phage clone produce the tissue factor fragment as a fusion protein with ..beta..-galactosidase. The isolated DNA fragment coding for part of the bovine tissue factor gene is estimated to be approximately 500 bases in length by agarose gel electrophoresis. The ..beta..-glactosidase - tissue factor fusion protein, subjected to Western immunoblotting, shows a protein product of approximately 130 Kd which is reactive to anti-tissue factor antibodies. This corresponds well to the 114 Kd ..beta..-galactosidase plus the predicted approx. 16 Kd fragment of tissue factor. Experiments are in progress to transfer the presumed tissue factor gene fragment into phage M13 for nucleotide sequence analysis.

  7. Accumulation of L-type Bovine Prions in Peripheral Nerve Tissues

    PubMed Central

    Imamura, Morikazu; Matsuura, Yuichi; Masujin, Kentaro; Shimizu, Yoshihisa; Shu, Yujing; Kurachi, Megumi; Kasai, Kazuo; Murayama, Yuichi; Fukuda, Shigeo; Onoe, Sadao; Hagiwara, Ken’ichi; Yamakawa, Yoshio; Sata, Tetsutaro; Mohri, Shirou; Okada, Hiroyuki; Yokoyama, Takashi

    2010-01-01

    We recently reported the intraspecies transmission of L-type atypical bovine spongiform encephalopathy (BSE). To clarify the peripheral pathogenesis of L-type BSE, we studied prion distribution in nerve and lymphoid tissues obtained from experimentally challenged cattle. As with classical BSE prions, L-type BSE prions accumulated in central and peripheral nerve tissues. PMID:20587193

  8. Real-time PCR for quantitative detection of bovine tissues in food and feed.

    PubMed

    Martín, Irene; García, Teresa; Fajardo, Violeta; Rojas, María; Hernández, Pablo E; González, Isabel; Martín, Rosario

    2008-03-01

    A real-time PCR approach with the SYBR Green detection system has been developed for the quantitative detection of bovine tissues in food and feedstuffs. The method combines the use of bovine-specific primers, which amplify an 84-bp fragment of the mitochondrial 12S rRNA gene, and universal primers, which amplify a 140-bp fragment of the nuclear 18S rRNA gene from eukaryotic DNA. The 18S rRNA primers are used as endogenous controls for the total content of PCR-amplifiable DNA in the sample. The specificity of the primers was tested against 18 animal species, including mammals, birds, and fish, as well as 6 plant species. Analysis of experimental bovine tissues-oats mixtures demonstrated the suitability of the assay for the detection of bovine DNA in mixtures containing as low as 0.1% of bovine tissues. The performance of the method is not affected by severe heat treatment (up to 133 degrees C for 20 min at 300 kPa). The reported PCR assay could be very useful for detecting bovine-derived ingredients in raw and heat-treated food and feedstuffs.

  9. Information Theoretical Analysis of a Bovine Gene Atlas Reveals Chromosomal Regions with Tissue Specific Gene Expression.

    USDA-ARS?s Scientific Manuscript database

    An essential step to understanding the genomic biology of any organism is to comprehensively survey its transcriptome. We present the Bovine Gene Atlas (BGA) a compendium of over 7.2 million unique 20 base Illumina DGE tags representing 100 tissue transcriptomes collected primarily from L1 Dominette...

  10. Light propagation along the pericardium meridian at human wrist as evidenced by the optical experiment and Monte Carlo method.

    PubMed

    Jiang, Yi-fan; Chen, Chang-shui; Liu, Xiao-mei; Liu, Rong-ting; Liu, Song-hao

    2015-04-01

    To explore the characteristics of light propagation along the Pericardium Meridian and its surrounding areas at human wrist by using optical experiment and Monte Carlo method. An experiment was carried out to obtain the distribution of diffuse light on Pericardium Meridian line and its surrounding areas at the wrist, and then a simplified model based on the anatomical structure was proposed to simulate the light transportation within the same area by using Monte Carlo method. The experimental results showed strong accordance with the Monte Carlo simulation that the light propagation along the Pericardium Meridian had an advantage over its surrounding areas at the wrist. The advantage of light transport along Pericardium Merdian line was related to components and structure of tissue, also the anatomical structure of the area that the Pericardium Meridian line runs.

  11. Residue depletion of eprinomectin in bovine tissues after subcutaneous administration.

    PubMed

    Jiang, Haiyang; Hou, Xiaolin; Ding, Shuangyang; Zhao, Sijun; He, Jihong; Shen, Jianzhong

    2005-11-16

    A study of the tissue depletion of eprinomectin (EPR) subcutaneously administered to cattle at a dose of 500 mg per kg of body weight was carried out. EPR concentrations were determined in muscle, liver, kidney, and fat. Twenty-four parasite-free cross cattle were treated with the EPR injectable oil formulation. Three treated animals (two males and one female) were selected randomly to be sacrificed at 1, 3, 7, 14, 21, 28, 42, and 56 days withdrawal after injection. EPR residue concentrations were determined using HPLC with fluorescence detection. Muscle samples showed the lowest EPR concentrations throughout the study period. The highest EPR concentrations at all sampling times were measured in liver tissue, indicating that liver is a target tissue for EPR. EPR concentrations in all of the tissues analyzed were below the accepted maximum residue limits recommended by the European Union at 8 days posttreatment.

  12. A catalogue of novel bovine long noncoding RNA across 18 tissues

    PubMed Central

    2015-01-01

    Long non-coding RNA (lncRNA) have been implicated in diverse biological roles including gene regulation and genomic imprinting. Identifying lncRNA in bovine across many differing tissue would contribute to the current repertoire of bovine lncRNA, and help further improve our understanding of the evolutionary importance and constraints of these transcripts. Additionally, it could aid in identifying sites in the genome outside of protein coding genes where mutations could contribute to variation in complex traits. This is particularly important in bovine as genomic predictions are increasingly used in genetic improvement for milk and meat production. Our aim was to identify and annotate novel long non coding RNA transcripts in the bovine genome captured from RNA Sequencing (RNA-Seq) data across 18 tissues, sampled in triplicate from a single cow. To address the main challenge in identifying lncRNA, namely distinguishing lncRNA transcripts from unannotated genes and protein coding genes, a lncRNA identification pipeline with a number of filtering steps was developed. A total of 9,778 transcripts passed the filtering pipeline. The bovine lncRNA catalogue includes MALAT1 and HOTAIR, both of which have been well described in human and mouse genomes. We attempted to validate the lncRNA in libraries from three additional cows. 726 (87.47%) liver and 1,668 (55.27%) blood class 3 lncRNA were validated with stranded liver and blood libraries respectively. Additionally, this study identified a large number of novel unknown transcripts in the bovine genome with high protein coding potential, illustrating a clear need for better annotations of protein coding genes. PMID:26496443

  13. A catalogue of novel bovine long noncoding RNA across 18 tissues.

    PubMed

    Koufariotis, Lambros T; Chen, Yi-Ping Phoebe; Chamberlain, Amanda; Vander Jagt, Christy; Hayes, Ben J

    2015-01-01

    Long non-coding RNA (lncRNA) have been implicated in diverse biological roles including gene regulation and genomic imprinting. Identifying lncRNA in bovine across many differing tissue would contribute to the current repertoire of bovine lncRNA, and help further improve our understanding of the evolutionary importance and constraints of these transcripts. Additionally, it could aid in identifying sites in the genome outside of protein coding genes where mutations could contribute to variation in complex traits. This is particularly important in bovine as genomic predictions are increasingly used in genetic improvement for milk and meat production. Our aim was to identify and annotate novel long non coding RNA transcripts in the bovine genome captured from RNA Sequencing (RNA-Seq) data across 18 tissues, sampled in triplicate from a single cow. To address the main challenge in identifying lncRNA, namely distinguishing lncRNA transcripts from unannotated genes and protein coding genes, a lncRNA identification pipeline with a number of filtering steps was developed. A total of 9,778 transcripts passed the filtering pipeline. The bovine lncRNA catalogue includes MALAT1 and HOTAIR, both of which have been well described in human and mouse genomes. We attempted to validate the lncRNA in libraries from three additional cows. 726 (87.47%) liver and 1,668 (55.27%) blood class 3 lncRNA were validated with stranded liver and blood libraries respectively. Additionally, this study identified a large number of novel unknown transcripts in the bovine genome with high protein coding potential, illustrating a clear need for better annotations of protein coding genes.

  14. Congenital absence of pericardium in babies with patent ductus arteriosus.

    PubMed

    Azhar, I O; Anas, R

    2013-06-01

    We report a case of two babies with absence of pericardium and patent ductus arteriosus (PDA). The absence of pericardium was found coincidentally during PDA ligation. The PDA was successfully ligated but the pericardium was not reconstructed. Postoperatively, the agenesis of the pericardium did not interfere with cardiac function.

  15. Visual and chemical tissue markers for bovine carcass components

    SciTech Connect

    Lary, R.Y.; Byers, F.M.; Cross, H.R.; Schelling, G.T.; Petersen, H.D.

    1988-04-01

    A two-component, nontoxic, quantifiable animal/carcass tracing system was developed using riboflavin as an on-premises, initial carcass identifier visible under longwave ultraviolet (UV) light and deuterium oxide (D/sub 2/O) as a tracer analytically quantified via fixed wavelength infrared spectrophotometry. Twenty-four cull cows and heifers were allocated into eight antemortem treatment groups (1, 12, 24, 48, 72, 96, 120, 144 h) for evaluation of the efficacy of riboflavin and D/sub 2/O as tissue tracers in postmortem meat tissues. All cattle were slaughtered using conventional procedures and inspection. To study postmortem riboflavin marker changes due to constant light exposure over time, fluorescence and emission intensity scores were obtained by a trained panel 24, 48, and 168 h postslaughter. The riboflavin marker intensity rating means for UV fluorescence were classified as identifiable on all carcasses when evaluated under UV light, but were classified as not identifiable when evaluated under ambient light. Deuterium oxide levels in all tissue water samples, regardless of antemortem infusion group, contained D/sub 2/O concentrations at least 2.5 times greater than those found in background water. Deuterium oxide was shown to disperse rapidly throughout living tissues. Correlations within animals for D/sub 2/O levels from blood and muscle were all highly significant (r = .99).

  16. An atlas of DNA methylation in diverse bovine tissues

    USDA-ARS?s Scientific Manuscript database

    We launched an effort to produce a reference cattle DNA methylation resource to improve animal production. We will employ experimental pipelines built around next generation sequencing technologies to map DNA methylation in cultured cells and primary tissues systems frequently involved in animal pro...

  17. Evaluation of mechanical behavior of soft tissue by means of random laser emission

    NASA Astrophysics Data System (ADS)

    Briones-Herrera, J. C.; Cuando-Espitia, N.; Sánchez-Arévalo, F. M.; Hernández-Cordero, J.

    2013-10-01

    We demonstrate the use of random laser emission for mechanical testing of bovine pericardium. An apparatus designed for tensile tests of soft and thin materials, incorporating optical and mechanical devices, allows for obtaining the mechanical behavior of the tissue samples. Using both, digital image correlation (DIC) and random laser emission analysis, the apparatus provides information regarding the response of the bovine pericardium under different stress levels. Our results show that changes in the spectral features of the random laser correlate well to the mechanical response obtained with conventional uniaxial tensile analysis coupled with DIC. Furthermore, parameters such as the shear and Young moduli are consistent with values reported previously and obtained with other techniques. Changes at the microstructural level of the tissue may thus be evaluated through spectral analysis of the random laser emission from biological samples.

  18. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Klaudia S G; Ramos, Carlos A N; Souza Filho, Antonio F; Vidal, Carlos E S; Vargas, Agueda P C; Roxo, Eliana; Rocha, Adalgiza S; Suffys, Philip N; Fonseca, Antônio A; Silva, Marcio R; Barbosa Neto, José D; Cerqueira, Valíria D; Araújo, Flábio R

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  19. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A.R.; Jorge, Klaudia S.G.; Ramos, Carlos A.N.; Souza Filho, Antonio F.; Vidal, Carlos E.S.; Vargas, Agueda P.C.; Roxo, Eliana; Rocha, Adalgiza S.; Suffys, Philip N.; Fonseca, Antônio A.; Silva, Marcio R.; Barbosa Neto, José D.; Cerqueira, Valíria D.; Araújo, Flábio R.

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:25242951

  20. Characterization of Transcriptional Complexity during Adipose Tissue Development in Bovines of Different Ages and Sexes

    PubMed Central

    Zhou, Yang; Sun, Jiajie; Li, Congjun; Wang, Yanhong; Li, Lan; Cai, Hanfang; Lan, Xianyong; Lei, Chuzhao; Zhao, Xin; Chen, Hong

    2014-01-01

    Background Adipose tissue has long been recognized to play an extremely important role in development. In bovines, it not only serves a fundamental function but also plays a key role in the quality of beef and, consequently, has drawn much public attention. Age and sex are two key factors that affect the development of adipose tissue, and there has not yet been a global study detailing the effects of these two factors on expressional differences of adipose tissues. Results In this study, total RNA from the back fat of fetal bovines, adult bulls, adult heifers and adult steers were used to construct libraries for Illumina next-generation sequencing. We detected the expression levels of 12,233 genes, with over 3,000 differently expressed genes when comparing fetal and adult patterns and an average of 1000 differently expressed genes when comparing adult patterns. Multiple Gene Ontology terms and pathways were found to be significantly enriched for these differentially expressed genes. Of the 12,233 detected genes, a total of 4,753 genes (38.85%) underwent alternative splicing events, and over 50% were specifically expressed in each library. Over 4,000 novel transcript units were discovered for one library, whereas only approximately 30% were considered to have coding ability, which supplied a large amount of information for the lncRNA study. Additionally, we detected 56,564 (fetal bovine), 65,154 (adult bull), 78,061 (adult heifer) and 86,965 (adult steer) putative single nucleotide polymorphisms located in coding regions of the four pooled libraries. Conclusion Here, we present, for the first time, a complete dataset involving the spatial and temporal transcriptome of bovine adipose tissue using RNA-seq. These data will facilitate the understanding of the effects of age and sex on the development of adipose tissue and supply essential information towards further studies on the genomes of beef cattle and other related mammals. PMID:24983926

  1. Experimental determination of constitutive equations for human and bovine brain tissue

    NASA Astrophysics Data System (ADS)

    Takhounts, Erik Grigorievich

    1998-09-01

    The purpose of this study was to determine experimentally the constitutive equations for brain tissue. Three series of experiments were performed in which the brain tissue was treated as a linear, quasi-linear and nonlinear isotropic viscoelastic material. Finite element analysis was performed and verified that simplifying assumptions made for developing constitutive equations were reasonable. Human and bovine brain samples were used to characterize linear behavior of brain tissue in the first series of tests. Single step tests with shear strains of up to 40% were performed to obtain stress-relaxation material functions for human and bovine brain tissue. The second series of experiments determined shear properties of bovine brain material by performing a set of single step loading stress-relaxation tests at the strain levels of up to 100%. For these tests, the theory of quasi-linear viscoelasticity (QLV) was employed to determine material properties. The third series of experiments involved nonlinear testing using single, two and three step loading stress-relaxation tests. The integral polynomial form of the third order Green-Rivlin constitutive equation was applied to model nonlinear behavior of the brain tissue. This representation describes the material behavior of brain tissue for the shear strains of up to 100%. The range of applicability for each viscoelastic theory was determined for brain material. It was found that for the strains of up to 40% a linear viscoelastic model is sufficient to describe material behavior. For the strains of up to 60% a quasi-linear model may be employed to describe the nonlinear behavior of brain tissue. At the strains of 60% and greater a time nonlinearity of brain material becomes significant and a nonlinear theory of viscoelasticity must be employed.

  2. Rapid analysis of aminoglycoside antibiotics in bovine tissues using disposable pipette extraction and ultrahigh performance liquid chromatography - tandem mass spectrometry

    USDA-ARS?s Scientific Manuscript database

    A high-throughput qualitative screening and identification method for 9 aminoglycosides of regulatory interest has been developed, validated, and implemented for bovine kidney, liver, and muscle tissues. The method involves extraction at previously validated conditions, cleanup using disposable pip...

  3. [Detection of bovine leukaemia virus (BLV) in tissue samples of naturally and experimentally infected cattle].

    PubMed

    Teifke, Jens P; Vahlenkamp, Thomas W

    2008-01-01

    Enzootic bovine leukaemia (EBL) which is caused by the bovine leukaemia virus (BLV) still plays a remarkable role despite a significant success in sanitation programmes. In the Federal Republic of Germany it was not possible to eradicate the disease until today. Sporadically during slaughter or necropsy of cattle neoplastic lesions of the lymphatic tissues are observed that need to be clarified with regard to BLV as etiological agent. Due to the fact that in most instances no serological data are available from the respective animals and blood drawings from the original holdings are not easy to obtain the polymerase chain reaction (PCR) opens new avenues as supplementary diagnostic tool to test unfixed lymphatic tissues for the presence of BLV proviral DNA. Lymph node tissues from 10 naturally or experimentally BLV-infected cattle, which have been monitored virologically and serologically, and tissues from 4 negative animals were processed, DNA was extracted and subjected to PCR to amplify BLV env gene specific sequences. The results show that in cattle with BLV-induced leukosis as well as in cattle, which were clinically healthy and unsuspicious at slaughter or at post-mortem, either with persistent lymphocytosis (PL) or without, BLV proviral DNA could be detected easily in samples of lymphatic tissues and in high concordance with serological data. In this article data from the National and OIE reference laboratory for EBL at the Friedrich-Loeffler-Institut (FLI, Germany) are presented. Elaborated laboratory protocols for processing of tissue samples and performing of BLV-PCR are recommended.

  4. A comparison of the fluorescence spectra of murine and bovine central nervous system (CNS) and other tissues

    USDA-ARS?s Scientific Manuscript database

    Here we describe a comparison of the fluorescence spectra of bovine tissues with murine tissues in order to determine whether spectral features are conserved and whether an appropriate and practical laboratory small animal model system could be identified to be used for investigation of tissue and a...

  5. Autofluorescence and Nonspecific Immunofluorescent Labeling in Frozen Bovine Intestinal Tissue Sections: Solutions for Multicolor Immunofluorescence Experiments.

    PubMed

    Jenvey, Caitlin J; Stabel, Judith R

    2017-09-01

    Autofluorescent compounds present in intestinal tissue often hinder the ability to utilize multiple, spectrally different, fluorophores. In addition, fixatives and blocking solutions may contribute to background autofluorescence or nonspecific immunofluorescent labeling. During immunofluorescence protocol development, autofluorescent pigments were observed in frozen bovine mid-ileal intestinal tissue sections. Coagulant fixatives, normal serum blocking, histochemical stains Sudan Black B (SBB) and 3,3'-diaminobenzidine (DAB), and spectral separation using imaging software were compared for their ability to reduce autofluorescence, as well as their effect on immunofluorescent labeling. Fluorescent pigments of frozen bovine mid-ileal intestinal tissue sections, most likely caused by eosinophils and lipofuscin, were masked successfully with a combination of DAB and SBB. Little to no statistical differences were observed for all other methods investigated; however, tissue fixed with 1:1 acetone methanol and 10% horse serum diluted in 0.05 M Tris buffer demonstrated lower mean fluorescence intensities. Spectral separation of specific immunofluorescent labeling from background autofluorescence is a simple method for removing unwanted fluorescence; however, successful separation is dependent on tissue and labeling quality.

  6. Viscoelastic properties of bovine orbital connective tissue and fat: constitutive models.

    PubMed

    Yoo, Lawrence; Gupta, Vijay; Lee, Choongyeop; Kavehpore, Pirouz; Demer, Joseph L

    2011-12-01

    Reported mechanical properties of orbital connective tissue and fat have been too sparse to model strain-stress relationships underlying biomechanical interactions in strabismus. We performed rheological tests to develop a multi-mode upper convected Maxwell (UCM) model of these tissues under shear loading. From 20 fresh bovine orbits, 30 samples of connective tissue were taken from rectus pulley regions and 30 samples of fatty tissues from the posterior orbit. Additional samples were defatted to determine connective tissue weight proportion, which was verified histologically. Mechanical testing in shear employed a triborheometer to perform: strain sweeps at 0.5-2.0 Hz; shear stress relaxation with 1% strain; viscometry at 0.01-0.5 s(-1) strain rate; and shear oscillation at 1% strain. Average connective tissue weight proportion was 98% for predominantly connective tissue and 76% for fatty tissue. Connective tissue specimens reached a long-term relaxation modulus of 668 Pa after 1,500 s, while corresponding values for fatty tissue specimens were 290 Pa and 1,100 s. Shear stress magnitude for connective tissue exceeded that of fatty tissue by five-fold. Based on these data, we developed a multi-mode UCM model with variable viscosities and time constants, and a damped hyperelastic response that accurately described measured properties of both connective and fatty tissues. Model parameters differed significantly between the two tissues. Viscoelastic properties of predominantly connective orbital tissues under shear loading differ markedly from properties of orbital fat, but both are accurately reflected using UCM models. These viscoelastic models will facilitate realistic global modeling of EOM behavior in binocular alignment and strabismus.

  7. Viscoelastic properties of bovine orbital connective tissue and fat: constitutive models

    PubMed Central

    Yoo, Lawrence; Gupta, Vijay; Lee, Choongyeop; Kavehpore, Pirouz

    2012-01-01

    Reported mechanical properties of orbital connective tissue and fat have been too sparse to model strain–stress relationships underlying biomechanical interactions in strabismus. We performed rheological tests to develop a multi-mode upper convected Maxwell (UCM) model of these tissues under shear loading. From 20 fresh bovine orbits, 30 samples of connective tissue were taken from rectus pulley regions and 30 samples of fatty tissues from the posterior orbit. Additional samples were defatted to determine connective tissue weight proportion, which was verified histologically. Mechanical testing in shear employed a triborheometer to perform: strain sweeps at 0.5–2.0 Hz; shear stress relaxation with 1% strain; viscometry at 0.01–0.5 s−1 strain rate; and shear oscillation at 1% strain. Average connective tissue weight proportion was 98% for predominantly connective tissue and 76% for fatty tissue. Connective tissue specimens reached a long-term relaxation modulus of 668 Pa after 1,500 s, while corresponding values for fatty tissue specimens were 290 Pa and 1,100 s. Shear stress magnitude for connective tissue exceeded that of fatty tissue by five-fold. Based on these data, we developed a multimode UCM model with variable viscosities and time constants, and a damped hyperelastic response that accurately described measured properties of both connective and fatty tissues. Model parameters differed significantly between the two tissues. Viscoelastic properties of predominantly connective orbital tissues under shear loading differ markedly from properties of orbital fat, but both are accurately reflected using UCM models. These viscoelastic models will facilitate realistic global modeling of EOM behavior in binocular alignment and strabismus. PMID:21207094

  8. Effects of trehalose supplementation on cell viability and oxidative stress variables in frozen-thawed bovine calf testicular tissue.

    PubMed

    Zhang, Xiao-Gang; Wang, Yan-Hua; Han, Cong; Hu, Shan; Wang, Li-Qiang; Hu, Jian-Hong

    2015-06-01

    Trehalose is widely used for cryopreservation of various cells and tissues. Until now, the effect of trehalose supplementation on cell viability and antioxidant enzyme activity in frozen-thawed bovine calf testicular tissue remains unexplored. The objective of the present study was to compare the effect of varying doses of trehalose in cryomedia on cell viability and key antioxidant enzymes activities in frozen-thawed bovine calf testicular tissue. Bovine calf testicular tissue samples were collected and cryopreserved in the cryomedias containing varying doses (0, 5, 10, 15, 20 and 25%; v/v) of trehalose, respectively. Cell viability, total antioxidant capacity (T-AOC) activity, catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione (GSH) content and malondialdehyde (MDA) content were measured and analyzed. The results showed that cell viability, T-AOC activity, SOD activity, CAT activity and GSH content of frozen-thawed bovine calf testicular tissue was decreased compared with that of fresh group (P<0.05). MDA content in frozen-thawed bovine calf testicular tissue was significantly increased compared with that of fresh group (P<0.05). The cryomedia added 15% trehalose exhibited the greatest percentage of cell viability and antioxidant enzyme activity (SOD and CAT) among frozen-thawed groups (P<0.05). Meanwhile, GSH content was the lowest among frozen-thawed groups (P<0.05). However, there were no significance differences in MDA content among the groups added 10, 15 and 20% trehalose (P>0.05). In conclusion, the cryomedia added 15% trehalose reduced the oxidative stress and improved the cryoprotective effect of bovine calf testicular tissue. Further studies are required to obtain more concrete results on the determination of antioxidant capacity of trehalose in frozen-thawed bovine calf testicular tissue.

  9. Relationship between streaming potential and compressive stress in bovine intervertebral tissue.

    PubMed

    Fujisaki, Kazuhiro; Tadano, Shigeru; Asano, Nozomu

    2011-09-02

    The intervertebral disc is formed by the nucleus pulposus (NP) and annulus fibrosus (AF), and intervertebral tissue contains a large amount of negatively charged proteoglycan. When this tissue becomes deformed, a streaming potential is induced by liquid flow with positive ions. The anisotropic property of the AF tissue is caused by the structural anisotropy of the solid phase and the liquid phase flowing into the tissue with the streaming potential. This study investigated the relationship between the streaming potential and applied stress in bovine intervertebral tissue while focusing on the anisotropy and loading location. Column-shaped specimens, 5.5 mm in diameter and 3 mm thick, were prepared from the tissue of the AF, NP and the annulus-nucleus transition region (AN). The loading direction of each specimen was oriented in the spinal axial direction, as well as in the circumferential and radial directions of the spine considering the anisotropic properties of the AF tissue. The streaming potential changed linearly with stress in all specimens. The linear coefficients k(e) of the relationship between stress and streaming potential depended on the extracted positions. These coefficients were not affected by the anisotropy of the AF tissue. In addition, these coefficients were lower in AF than in NP specimens. Except in the NP specimen, the k(e) values were higher under faster compression rate conditions. In cyclic compression loading the streaming potential changed linearly with compressive stress, regardless of differences in the tissue and load frequency.

  10. Investigation of the suitability of decellularized porcine pericardium in mitral valve reconstruction.

    PubMed

    Morticelli, Lucrezia; Thomas, Daniel; Roberts, Nicholas; Ingham, Eileen; Korossis, Sotirios

    2013-05-01

    Autologous and glutaraldehyde-treated xenogeneic and homogeneic pericardium has been used extensively in mitral valve repair, but there are a number of limitations associated with its use. These include calcification, limited durability and lack of in vivo regeneration with glutaraldehyde-treated xenografts, as well as the sacrifice of the patient's own pericardium in the case of repair with autologous pericardium. The study aim was to investigate the suitability of decellularized porcine pericardium for heterotopic repair of the mitral valve leaflets, and its potential to regenerate through endogenous cell repopulation in vivo, or in vitro cell seeding prior to implantation. Fresh porcine anterior and posterior mitral valve leaflets, together with fresh and decellularized porcine pericardium, were tested histologically, biochemically and biomechanically to investigate potential similarities and differences between the different types of tissue. Subsequently, the decellularized pericardial scaffolds were tested both in terms of biocompatibility, using contact and extract cytotoxicity assays, and in terms of regenerative capacity through porcine mesenchymal stem cell (pMSC) seeding. Histological examination of fresh pericardium and leaflets showed the typical trilaminar and quadlaminar structures of the two tissues, respectively. No cell remnants were observed in the decellularized pericardium, whereas the histoarchitecture of the collagen, elastin and glycosaminoglycan (GAG) matrix appeared well preserved. Significant differences were found in the GAG and hydroxyproline contents and the biomechanics between the leaflet and the pericardial groups. No indication of cytotoxicity was observed with the decellularized pericardial scaffolds. The optimum cell seeding density of pMSCs was 1 x 10(5) cells per cm2, which represented the lowest density at which the cells were capable of repopulating the scaffold by migrating through its full thickness. Porcine mitral valve

  11. Rapid tissue dissolution efficiency of electrically-activated sodium hypochlorite on bovine muscle

    PubMed Central

    Ertugrul, Ihsan Furkan; Maden, Murat; Orhan, Ekim Onur; Ozkorucuklu, Sabriye Percin; Aglarca, Ali Vasfi

    2014-01-01

    Objective: Sodium hypochlorite (NaOCl) is a common antimicrobial and tissue-dissolving irrigant. The aim of this in vitro study is to evaluate and compare dissolution capacities of sodium hypochlorite solutions after electrically activation (E-NaOCl) on bovine muscle specimens at various time periods and concentrations. Materials and Methods: Three sodium hypochlorite solutions of 1.25%, 2.5%, and 5% were tested at 3-min. and 5-min. with and without activation by electrically. Distilled water and NaOCl solutions without electrically activation were used as controls. Pieces of bovine muscle tissue (34 ± 2 mg) were placed in 10 mL of each solution at room temperature. In the group of E-NaOCl, electrically activation was performed through the potentiostat. The tissue specimens were weighed before and after treatment, and the percentage of weight loss was calculated. Results: Weight loss of the tissue increased with the concentration of E-NaOCl and NaOCl. Higher concentration and electrically activation considerably enhanced the efficacy of sodium hypochlorite. The effect of electrically activation on tissue dissolution was much greater than that of same concentrations in the groups of NaOCl (P < 0.001). Tissue weight loss was significantly higher in 2.5% and 5% E-NaOCl at 3 min. than in 2.5% and 5% NaOCl at 5 min. (P < 0.05). There were not any significant differences between the 2.5% E-NaOCl and 5% NaOCl at 5 min. (P > 0.05). Conclusion: Electrically activation can improve the tissue-dissolving effectiveness of sodium hypochlorite. PMID:25512725

  12. Rapid tissue dissolution efficiency of electrically-activated sodium hypochlorite on bovine muscle.

    PubMed

    Ertugrul, Ihsan Furkan; Maden, Murat; Orhan, Ekim Onur; Ozkorucuklu, Sabriye Percin; Aglarca, Ali Vasfi

    2014-10-01

    Sodium hypochlorite (NaOCl) is a common antimicrobial and tissue-dissolving irrigant. The aim of this in vitro study is to evaluate and compare dissolution capacities of sodium hypochlorite solutions after electrically activation (E-NaOCl) on bovine muscle specimens at various time periods and concentrations. Three sodium hypochlorite solutions of 1.25%, 2.5%, and 5% were tested at 3-min. and 5-min. with and without activation by electrically. Distilled water and NaOCl solutions without electrically activation were used as controls. Pieces of bovine muscle tissue (34 ± 2 mg) were placed in 10 mL of each solution at room temperature. In the group of E-NaOCl, electrically activation was performed through the potentiostat. The tissue specimens were weighed before and after treatment, and the percentage of weight loss was calculated. Weight loss of the tissue increased with the concentration of E-NaOCl and NaOCl. Higher concentration and electrically activation considerably enhanced the efficacy of sodium hypochlorite. The effect of electrically activation on tissue dissolution was much greater than that of same concentrations in the groups of NaOCl (P < 0.001). Tissue weight loss was significantly higher in 2.5% and 5% E-NaOCl at 3 min. than in 2.5% and 5% NaOCl at 5 min. (P < 0.05). There were not any significant differences between the 2.5% E-NaOCl and 5% NaOCl at 5 min. (P > 0.05). Electrically activation can improve the tissue-dissolving effectiveness of sodium hypochlorite.

  13. Outcomes of shunt tube coverage with glycerol preserved cornea versus pericardium.

    PubMed

    Wigton, Eric; C Swanner, Jason; Joiner, Wade; Feldman, Alex; McGwin, Gerald; Huisingh, Carrie; Curcio, Christine A; Girkin, Christopher A

    2014-01-01

    Pericardium is a biomaterial widely used for covering the outflow tubes of glaucoma drainage devices. Recently, glycerol preserved cornea has been introduced as an alternative that offers durability and improved cosmesis because of its clarity. We retrospectively reviewed 262 patients in the University of Alabama Birmingham Glaucoma Service who underwent shunt procedures using either cornea tissue or pericardium to cover the tube. The primary outcome measure was the number of erosions of the covering material. Nine out of 101 (8.9%) patients in the pericardium covered group experienced an erosion compared with 3 out of 161 (1.9%) in the cornea covered group. A significant difference was reached with P=0.0125. Median follow-up was 440 days for the cornea group and 331 days for the pericardium group. The type of glaucoma (primary open-angle glaucoma vs. secondary glaucoma) was not associated with the risk of erosion (odds ratio, 0.501; 95% confidence interval, 0.204-1.234). The median time to exposure was 252 days in the pericardium group and 440 days in the cornea group (P=0.0017).

  14. Multicystic mesothelioma of the pericardium.

    PubMed

    Morita, Shigeki; Goto, Akiteru; Sakatani, Takashi; Ota, Satoshi; Murakawa, Tomohiro; Nakajima, Jun; Maeda, Eriko; Fukayama, Masashi

    2011-05-01

    Multicystic mesothelioma is a well recognized but rare serosal tumor which mainly arises from the peritoneum in women and is considered as a benign lesion. This is the second case report of pericardial multicystic mesothelioma, which took a fatal clinical course. A 63-year-old man presented with pitting edema, shortness of breath, and hoarseness. Radiological investigations revealed solid and cystic tumor of the pericardium which was continuously extending into the mediastinum and the liver. Pericardial biopsy showed micro-cystic tumor lined by single layer of mesothelial cells without atypia, and the diagnosis was multicystic mesothelioma. Curative surgery could not be performed, and three years and four months later, the patient died because of the direct compression of the heart by the tumor. At autopsy, the tumor was found to be directly extending into the right pleural cavity and the right lung, besides the mediastinum and the liver. Neither malignant transformation nor metastatic tumor was identified. © 2011 The Authors. Pathology International © 2011 Japanese Society of Pathology and Blackwell Publishing Asia Pty Ltd.

  15. Histopathology and immunohistochemistry of tissues outside central nervous system in bovine rabies.

    PubMed

    Abreu, Camila C; Nakayama, Priscilla A; Nogueira, Clayton I; Mesquita, Leonardo P; Lopes, Priscila F R; Wouters, Flademir; Varaschin, Mary S; Bezerra, Pedro S

    2014-08-01

    We performed a histopathological and immunohistochemical study of tissues outside the central nervous system in 48 cases of bovine rabies confirmed by direct immunofluorescence and/or immunohistochemistry (IHC) of the central nervous system. In the bovines of this study, mononuclear inflammation in all ganglia (trigeminal, spinal, stellate, and celiac) and adrenal medulla was observed. This injury also occurred in 85 % of neuro-pituitaries in 55 % of pars intermediate and 15 % of the pars distalis of pituitary evaluated. IHC was positive in 92.31 % of lumbar spinal ganglia, 90.9 % of trigeminal ganglia, stellate ganglia of 41.67 and 16.67 % of the celiac ganglia. One of the evaluated adrenal (1/17) showed strong immunohistochemical labeling in the cytoplasm of pheochromocytes. The pituitary IHC was positive in one case in the neurohypophysis (1/20) and in one case in the pars intermedia of the adenohypophysis (1/20). Data from this study indicate that in suspected cases of rabies, besides the complex pituitary rete mirabile and trigeminal ganglion, the evaluation of other ganglia, particularly the lumbar spinal, and adrenal may also contribute to the diagnosis and understanding of the clinical presentation and pathogenesis of the disease in bovines.

  16. Distribution of G/sub o. cap alpha. / mRNA and protein in bovine tissues

    SciTech Connect

    Price, S.R.; Tsai, S.C.; Adamik, R.; Angus, C.W.; Van Meurs, K.P.; Czarnecki, S.; Bruckwick, E.C.; Moss, J.; Vaughan, M.

    1987-05-01

    G/sub o..cap alpha../ is a 39 kDa guanyl nucleotide-binding protein similar in structure and function to G/sub s..cap alpha../ and G/sub i..cap alpha../ in the adenylate cyclase complex and transducin (G/sub t..cap alpha../) in the retinal photon receptor system. A bovine retinal cDNA clone, lambdaG09, that encodes the complete amino acid sequence of G/sub o..cap alpha../ has been isolated. Nick-translated lambdaG09 cDNA and a 5' end-labeled oligonucleotide probe complementary to a 24 base sequence unique to G/sub o..cap alpha../ were used as probes for Northern analysis of poly(A)/sup +/ RNA from bovine tissues. A major 4.0 kb mRNA was detected in brain and retina and in lesser amounts in heart. Several smaller mRNAs also hybridized with both probes in these tissues and in liver and lung. G/sub o..cap alpha../ protein was identified using rabbit polyclonal antibodies directed against purified bovine G/sub o..cap alpha../ and pertussis toxin-catalyzed (/sup 32/P)ADP-ribosylation. Soluble and membrane proteins were incubated with toxin and (/sup 32/P)NAD and then separated by gel electrophoresis before transfer to nitrocellulose for immunoreaction and subsequent autoradiography. A radiolabeled and immunoreactive 39 kDa membrane protein was found principally in retina and brain, and to a lesser extent, in heart. Thus, in the tissues examined, distribution of the 4.0 kb mRNA parallels that of the immunoreactive G/sub o..cap alpha../ with relatively small amounts in heart and larger amounts in brain and retina.

  17. Enteric, hepatic and muscle tissue development of goat kids fed with lyophilized bovine colostrum.

    PubMed

    Moretti, D B; Nordi, W M; Lima, A L; Pauletti, P; Machado-Neto, R

    2014-04-01

    The objective of this study was to investigate the development of the enteric, hepatic and muscle tissues in goat kids fed with lyophilized bovine colostrum in the transition period of passive immunity to early active immunity. At 0, 7 and 14 h of life, 15 male newborns received 5% of their body weight of lyophilized bovine colostrum and 14 male newborns received goat colostrum, both with 55 mg/ml of IgG. Samples of the duodenum, jejunum, ileum, liver and muscle were collected at 18, 36 and 96 h of life to quantify total protein, DNA and RNA contents. In the jejunum and ileum, the highest levels of total protein and higher protein/RNA ratio were observed at 18 h (p < 0.05). There were no differences in DNA contents in any intestinal segment (p > 0.05). At 96 h, maximum levels of RNA were observed in the jejunum and ileum (p < 0.05) and higher RNA/DNA ratio in the three intestinal segments (p < 0.05), showing increased ability to synthesize intracellular RNA and proteins. The LBC group showed higher protein content and higher protein/DNA and protein/RNA ratios in the jejunum, a higher DNA content in the liver (p < 0.05) and a higher protein/RNA ratio in the muscle tissue (p < 0.05). In the muscle, higher protein and DNA levels were also found at 96 h (p < 0.05). Indicators of cellular activity suggest greater absorption of proteins from lyophilized bovine colostrum and increased cell maturity in the enteric and muscle tissues in the first hours of goat kids' life.

  18. Transcriptomic comparison of primary bovine horn core carcinoma culture and parental tissue at early stage

    PubMed Central

    Shil, Sharadindu; Joshi, R. S.; Joshi, C. G.; Patel, A. K.; Shah, Ravi K.; Patel, Namrata; Jakhesara, Subhash J.; Kundu, Sumana; Reddy, Bhaskar; Koringa, P. G.; Rank, D. N.

    2017-01-01

    Aim: Squamous cell carcinoma or SCC of horn in bovines (bovine horn core carcinoma) frequently observed in Bos indicus affecting almost 1% of cattle population. Freshly isolated primary epithelial cells may be closely related to the malignant epithelial cells of the tumor. Comparison of gene expression in between horn’s SCC tissue and its early passage primary culture using next generation sequencing was the aim of this study. Materials and Methods: Whole transcriptome sequencing of horn’s SCC tissue and its early passage cells using Ion Torrent PGM were done. Comparative expression and analysis of different genes and pathways related to cancer and biological processes associated with malignancy, proliferating capacity, differentiation, apoptosis, senescence, adhesion, cohesion, migration, invasion, angiogenesis, and metabolic pathways were identified. Results: Up-regulated genes in SCC of horn’s early passage cells were involved in transporter activity, catalytic activity, nucleic acid binding transcription factor activity, biogenesis, cellular processes, biological regulation and localization and the down-regulated genes mainly were involved in focal adhesion, extracellular matrix receptor interaction and spliceosome activity. Conclusion: The experiment revealed similar transcriptomic nature of horn’s SCC tissue and its early passage cells. PMID:28246447

  19. Detection of bovine central nervous system tissue as bovine spongiform encephalopathy risk material by real-time reverse transcriptase-PCR in raw and cooked beef products.

    PubMed

    Shi, Xi-Ju; Ma, Gui-Ping; Li, Bing-Ling; Yang, Jin-Liang; Yu-Wang; Li, Yan-Xin; Liu, Xu-Hui; Liu, Quan-Guo

    2008-01-01

    There is increasing evidence of the association of the new variant of Creutzfeldt-Jacob disease (nvCJD) in humans with bovine spongiform encephalopathy (BSE) in cattle. Many countries established legislation of banning central nervous system (CNS) tissues, which are regarded as BSE-specified risk materials (SRM), in human food supply because of the potential transmission of BSE to humans. A real-time reverse transcriptase-PCR assay using the bovine glial fibrillary acidic protein (GFAP) mRNA template for the detection of CNS tissues in raw and cooked beef products was developed in this study. The results showed that (1) this method can detect CNS tissues from bovine and ovine origins, but not from porcine and avian ones; (2) GFAP mRNA can only be detected from brain and spinal cords rather than other tissues; (3) the GFAP mRNA was detectable in CNS tissues even after dilution to 0.001%; and (4) the assay was unaffected by heat treatment at 100 degrees C for 30 min or storage at room temperature for 4 days, and at 4 degrees C for at least 15 days.

  20. Mechanical compliance and immunological compatibility of fixative-free decellularized/cryopreserved human pericardium.

    PubMed

    Vinci, Maria Cristina; Tessitore, Giulio; Castiglioni, Laura; Prandi, Francesca; Soncini, Monica; Santoro, Rosaria; Consolo, Filippo; Colazzo, Francesca; Micheli, Barbara; Sironi, Luigi; Polvani, Gianluca; Pesce, Maurizio

    2013-01-01

    The pericardial tissue is commonly used to produce bio-prosthetic cardiac valves and patches in cardiac surgery. The procedures adopted to prepare this tissue consist in treatment with aldehydes, which do not prevent post-graft tissue calcification due to incomplete xeno-antigens removal. The adoption of fixative-free decellularization protocols has been therefore suggested to overcome this limitation. Although promising, the decellularized pericardium has not yet used in clinics, due to the absence of proofs indicating that the decellularization and cryopreservation procedures can effectively preserve the mechanical properties and the immunologic compatibility of the tissue. The aim of the present work was to validate a procedure to prepare decellularized/cryopreserved human pericardium which may be implemented into cardiovascular homograft tissue Banks. The method employed to decellularize the tissue completely removed the cells without affecting ECM structure; furthermore, uniaxial tensile loading tests revealed an equivalent resistance of the decellularized tissue to strain, before and after the cryopreservation, in comparison with the fresh tissue. Finally, immunological compatibility, showed a minimized host immune cells invasion and low levels of systemic inflammation, as assessed by tissue transplantation into immune-competent mice. Our results indicate, for the first time, that fixative-free decellularized pericardium from cadaveric tissue donors can be banked according to Tissue Repository-approved procedures without compromising its mechanical properties and immunological tolerance. This tissue can be therefore treated as a safe homograft for cardiac surgery.

  1. Mechanical Compliance and Immunological Compatibility of Fixative-Free Decellularized/Cryopreserved Human Pericardium

    PubMed Central

    Vinci, Maria Cristina; Tessitore, Giulio; Castiglioni, Laura; Prandi, Francesca; Soncini, Monica; Santoro, Rosaria; Consolo, Filippo; Colazzo, Francesca; Micheli, Barbara; Sironi, Luigi; Polvani, Gianluca; Pesce, Maurizio

    2013-01-01

    Background The pericardial tissue is commonly used to produce bio-prosthetic cardiac valves and patches in cardiac surgery. The procedures adopted to prepare this tissue consist in treatment with aldehydes, which do not prevent post-graft tissue calcification due to incomplete xeno-antigens removal. The adoption of fixative-free decellularization protocols has been therefore suggested to overcome this limitation. Although promising, the decellularized pericardium has not yet used in clinics, due to the absence of proofs indicating that the decellularization and cryopreservation procedures can effectively preserve the mechanical properties and the immunologic compatibility of the tissue. Principal Findings The aim of the present work was to validate a procedure to prepare decellularized/cryopreserved human pericardium which may be implemented into cardiovascular homograft tissue Banks. The method employed to decellularize the tissue completely removed the cells without affecting ECM structure; furthermore, uniaxial tensile loading tests revealed an equivalent resistance of the decellularized tissue to strain, before and after the cryopreservation, in comparison with the fresh tissue. Finally, immunological compatibility, showed a minimized host immune cells invasion and low levels of systemic inflammation, as assessed by tissue transplantation into immune-competent mice. Conclusions Our results indicate, for the first time, that fixative-free decellularized pericardium from cadaveric tissue donors can be banked according to Tissue Repository-approved procedures without compromising its mechanical properties and immunological tolerance. This tissue can be therefore treated as a safe homograft for cardiac surgery. PMID:23705010

  2. Congenital complete and partial absence of the left pericardium.

    PubMed

    Jurko, Alexander; Minarik, Milan; Cisarikova, Viera; Polacek, Hubert; Schusterova, Ingrid

    2013-09-01

    Congenital absence of pericardium is a rare malformation. We report 2 young patients with a diagnosis of congenital absence of the pericardium. The posteroanterior view of the chest X-ray showed displacement of the left cardiac border into the left hemithorax. Unusual acoustical windows and abnormal cardiac and septal motion during echocardiography suggested the diagnosis of congenital absence of pericardium. Magnetic resonance imaging definitive confirmed diagnosis of congenital absence of pericardium.

  3. Thermomechanical analysis of collagen crosslinking in the developing lamb pericardium.

    PubMed

    Naimark, W A; Waldman, S D; Anderson, R J; Suzuki, B; Pereira, C A; Lee, J M

    1998-01-01

    In a developing lamb model, we have used hydrothermal isometric tension (HIT) techniques to assess collagen crosslink stability and its contribution to the mechanical properties of the pericardium. Strip samples of tissue were either: (i) heated to a 90 degrees C isotherm or (ii) heated in 5 degrees C increments between 60-90 degrees C and then 93 and 97 degrees C isotherms. The half-life of stress relaxation associated with peptide bond hydrolysis (t1/2) was calculated at each isotherm. The activation energy, Eact, for the hydrolysis-associated relaxation process was also calculated using the data from the stepwise HIT tests--a technical improvement which significantly reduces the experimental time required to develop statistically valid measurements. Crosslinking in the pericardium increased during development and was demonstrated both by thermoelasticity and by resistance to enzymatic solubilization. We observed greater conversation to thermally stable crosslinks upon maturation, the ratio of the NaBH4-stabilized/unstabilized half-lives peaking at 21 days postnatal. Whereas tissue from lambs (119 day fetal, and 3 day and 21 day postpartum) showed an early maximum and rapid decay of force, NaBH4 stabilization significantly increased thermal stability and yielded profiles similar to those in adult tissue.

  4. Reinforced Pericardium as a Hybrid Material for Cardiovascular Applications

    PubMed Central

    Bracaglia, Laura G.; Yu, Li; Hibino, Narutoshi

    2014-01-01

    Pericardium-based cardiovascular devices are currently bound by a 10-year maximum lifetime due to detrimental calcification and degradation. The goal of this work is to develop a novel synthetic material to create a lasting replacement for malfunctioning or diseased tissue in the cardiovascular system. This study couples poly(propylene fumarate) (PPF) and a natural biomaterial together in an unprecedented hybrid composite and evaluates the composite versus the standard glutaraldehyde-treated tissue. The polymer reinforcement is hypothesized to provide initial physical protection from proteolytic enzymes and degradation, but leave the original collagen and elastin matrix unaltered. The calcification rate and durability of the hybrid material are evaluated in vitro and in an in vivo subdermal animal model. Results demonstrate that PPF is an effective support and leads to significantly less calcium deposition, important metrics when evaluating cardiovascular material. By avoiding chemical crosslinking of the tissue and associated side effects, PPF-reinforced pericardium as a biohybrid material offers a promising potential direction for further development in cardiovascular material alternatives. Eliminating the basis for the majority of cardiovascular prosthetic failures could revolutionize expectations for extent of cardiovascular repair. PMID:25236439

  5. Reinforced pericardium as a hybrid material for cardiovascular applications.

    PubMed

    Bracaglia, Laura G; Yu, Li; Hibino, Narutoshi; Fisher, John P

    2014-11-01

    Pericardium-based cardiovascular devices are currently bound by a 10-year maximum lifetime due to detrimental calcification and degradation. The goal of this work is to develop a novel synthetic material to create a lasting replacement for malfunctioning or diseased tissue in the cardiovascular system. This study couples poly(propylene fumarate) (PPF) and a natural biomaterial together in an unprecedented hybrid composite and evaluates the composite versus the standard glutaraldehyde-treated tissue. The polymer reinforcement is hypothesized to provide initial physical protection from proteolytic enzymes and degradation, but leave the original collagen and elastin matrix unaltered. The calcification rate and durability of the hybrid material are evaluated in vitro and in an in vivo subdermal animal model. Results demonstrate that PPF is an effective support and leads to significantly less calcium deposition, important metrics when evaluating cardiovascular material. By avoiding chemical crosslinking of the tissue and associated side effects, PPF-reinforced pericardium as a biohybrid material offers a promising potential direction for further development in cardiovascular material alternatives. Eliminating the basis for the majority of cardiovascular prosthetic failures could revolutionize expectations for extent of cardiovascular repair.

  6. Microwave Ablation Compared with Radiofrequency Ablation for Breast Tissue in an Ex Vivo Bovine Udder Model

    SciTech Connect

    Tanaka, Toshihiro; Westphal, Saskia; Isfort, Peter; Braunschweig, Till; Penzkofer, Tobias Bruners, Philipp; Kichikawa, Kimihiko; Schmitz-Rode, Thomas Mahnken, Andreas H.

    2012-08-15

    Purpose: To compare the effectiveness of microwave (MW) ablation with radiofrequency (RF) ablation for treating breast tissue in a nonperfused ex vivo model of healthy bovine udder tissue. Materials and Methods: MW ablations were performed at power outputs of 25W, 35W, and 45W using a 915-MHz frequency generator and a 2-cm active tip antenna. RF ablations were performed with a bipolar RF system with 2- and 3-cm active tip electrodes. Tissue temperatures were continuously monitored during ablation. Results: The mean short-axis diameters of the coagulation zones were 1.34 {+-} 0.14, 1.45 {+-} 0.13, and 1.74 {+-} 0.11 cm for MW ablation at outputs of 25W, 35W, and 45W. For RF ablation, the corresponding values were 1.16 {+-} 0.09 and 1.26 {+-} 0.14 cm with electrodes having 2- and 3-cm active tips, respectively. The mean coagulation volumes were 2.27 {+-} 0.65, 2.85 {+-} 0.72, and 4.45 {+-} 0.47 cm{sup 3} for MW ablation at outputs of 25W, 35W, and 45W and 1.18 {+-} 0.30 and 2.29 {+-} 0.55 cm{sup 3} got RF ablation with 2- and 3-cm electrodes, respectively. MW ablations at 35W and 45W achieved significantly longer short-axis diameters than RF ablations (P < 0.05). The highest tissue temperature was achieved with MW ablation at 45W (P < 0.05). On histological examination, the extent of the ablation zone in MW ablations was less affected by tissue heterogeneity than that in RF ablations. Conclusion: MW ablation appears to be advantageous with respect to the volume of ablation and the shape of the margin of necrosis compared with RF ablation in an ex vivo bovine udder.

  7. Angiosarcoma of the pericardium: a case report.

    PubMed

    Ma, Guo-Tao; Liu, Jian-Zhou; Miao, Qi; Cui, Quan-Cai

    2015-01-01

    Primary malignant tumors of the pericardium are rare, and most primary malignant pericardium tumors are mesotheliomas. Primary pericardial angiosarcoma is extremely rare, and it is associated with a poor prognosis. We report of a 47-year-old woman who complained of activity-related chest tightness and shortness of breath. Computed tomography, magnetic resonance imaging, and transesophageal echocardiography revealed an enlarged pericardium with hematic and solid components. An exploratory pericardiotomy was performed, and the results of the histological examination were suggestive of spindle cell hemangioendothelioma. She survived for 9 months after surgery without chemotherapy and radiotherapy, and she had a relatively good quality of life. Primary pericardial angiosarcoma is difficult to diagnose, and it has a poor prognosis. Pericardiotomy, radiation therapy, and chemotherapy were associated with a prolongation of survival.

  8. Tissue distribution of bovine viral diarrhea virus antigens in persistently infected cattle.

    PubMed

    Shin, T; Acland, H

    2001-08-01

    The tissue distribution and cellular localization of viral antigens in three cattle with persistent bovine viral diarrhea virus (BVDV) infection was studied. In three cases, necropsy findings of oral ulcers, abmasal ulcers and necrosis of Peyer's patches were suspected have been caused by BVDV infection. Non-cytopathic BVDV was isolated from a tissue pool of liver, kidneys and spleen. Immunohistochemical detection of BVDV showed that BVDV antigens were detected in both epithelial and nonepithelial cells in all examined organs, including the gastrointestinal tract, liver, pancreas, lung, lymphatic organs (spleen, lymph nodes), adrenal gland, ovary, uterus, and the mammary gland. These findings support the hypothesis that animals with persistent BVDV infection spread BVDV through all routes, and that infertility in BVDV infection is associated with the infection of BVDV in the ovaries and uteri.

  9. Effects of pulsed mid-IR lasers on bovine knee joint tissues

    NASA Astrophysics Data System (ADS)

    Vari, Sandor G.; Shi, Wei-Qiang; Pergadia, Vani R.; Duffy, J. T.; Miller, J. M.; van der Veen, Maurits J.; Weiss, Andrew B.; Fishbein, Michael C.; Grundfest, Warren S.

    1993-07-01

    We investigated the effect of varying Tm:YAG (2.014 micrometers ) and Ho:YAG (2.130 micrometers ) laser parameters on ablation rate and consequent thermal damage. Mid-infrared wavelengths are strongly absorbed by most biological tissues due to the tissue's high water content. The ablation rate of fresh bovine knee joint tissues (fibrous cartilage, hyaline cartilage, and bone) in saline was assessed as a function of radiant exposure (160 - 950 J/cm2), at pulse widths of 200 microsecond(s) ec for Tm:YAG and 250 microsecond(s) ec for Ho:YAG and a repetition rate of 2 Hz. All tissues used in this study could be efficiently ablated using two micron lasers. The mechanism of action is likely related to the formation and collapse of cavitation bubbles, associated with mid-infrared lasers. We concluded that the Tm:YAG and Ho:YAG lasers are capable of effective knee joint tissue ablation.

  10. Morphological analysis of the hagfish heart. II. The venous pole and the pericardium.

    PubMed

    Icardo, José M; Colvee, Elvira; Schorno, Sarah; Lauriano, Eugenia R; Fudge, Douglas S; Glover, Chris N; Zaccone, Giacomo

    2016-07-01

    The morphological characteristics of the venous pole and pericardium of the heart were examined in three hagfish species, Myxine glutinosa, Eptatretus stoutii, and Eptatretus cirrhatus. In these species, the atrioventricular (AV) canal is long, funnel-shaped and contains small amounts of myocardium. The AV valve is formed by two pocket-like leaflets that lack a papillary system. The atrial wall is formed by interconnected muscle trabeculae and a well-defined collagenous system. The sinus venosus (SV) shows a collagenous wall and is connected to the left side of the atrium. An abrupt collagen-muscle boundary marks the SV-atrium transition. It is hypothesized that the SV is not homologous to that of other vertebrates which could have important implications for understanding heart evolution. In M. glutinosa and E. stoutii, the pericardium is a closed bag that hangs from the tissues dorsal to the heart and encloses both the heart and the ventral aorta. In contrast, the pericardium is continuous with the loose periaortic tissue in E. cirrhatus. In all three species, the pericardium ends at the level of the SV excluding most of the atrium from the pericardial cavity. In M. glutinosa and E. stoutii, connective bridges extend between the base of the aorta and the ventricular wall. In E. cirrhatus, the connections between the periaortic tissue and the ventricle may carry blood vessels that reach the ventricular base. A further difference specific to E. cirrhatus is that the adipose tissue associated with the pericardium contains thyroid follicles. J. Morphol. 277:853-865, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  11. Congenital defects of the pericardium: a review.

    PubMed

    Shah, Ankit B; Kronzon, Itzhak

    2015-08-01

    Pericardial defects are a rare disorder that can be characterized as acquired or congenital. Congenital defects can be further characterized by location and size of the defect, e.g. left- or right-sided and partial or complete absence of the pericardium. While physical examination and electrocardiogram are not diagnostic, chest radiographs and echocardiography have findings that should alert the clinician to the absence of the pericardium as a possible diagnosis. Despite its limitations with visualizing the normal pericardium in areas of minimal adipose, cardiac magnetic resonance is currently the gold standard for diagnosing the congenital absence of the pericardium. Patients have a similar life expectancy to those without pericardial defects; however in certain cases, herniation and strangulation of cardiac chambers can be life threatening and lead to sudden cardiac death. Treatment is tailored to the patient's symptoms, presentation, and the size and location of the defect. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email: journals.permissions@oup.com.

  12. The behavior of cardiac progenitor cells on macroporous pericardium-derived scaffolds.

    PubMed

    Rajabi-Zeleti, Sareh; Jalili-Firoozinezhad, Sasan; Azarnia, Mahnaz; Khayyatan, Fahimeh; Vahdat, Sadaf; Nikeghbalian, Saman; Khademhosseini, Ali; Baharvand, Hossein; Aghdami, Nasser

    2014-01-01

    Cardiovascular diseases hold the highest mortality rate among other illnesses which reveals the significance of current limitations in common therapies. Three-dimensional (3D) scaffolds have been utilized as potential therapies for treating heart failure following myocardial infarction (MI). In particular, native tissues have numerous properties that make them potentially useful scaffolding materials for recreating the native cardiac extracellular matrix (ECM). Here, we have developed a pericardium-derived scaffold that mimics the natural myocardial extracellular environment and investigated its properties for cardiac tissue engineering. Human pericardium membranes (PMs) were decellularized to yield 3D macroporous pericardium scaffolds (PSs) with well-defined architecture and interconnected pores. PSs enabled human Sca-1(+) cardiac progenitor cells (CPCs) to migrate, survive, proliferate and differentiate at higher rates compared with decellularized pericardium membranes (DPMs) and collagen scaffolds (COLs). Interestingly, histological examination of subcutaneous transplanted scaffolds after one month revealed low immunological response, enhanced angiogenesis and cardiomyocyte differentiation in PSs compared to DPMs and COLs. This research demonstrates the feasibility of fabricating 3D porous scaffolds from native ECMs and suggests the therapeutic potential of CPC-seeded PSs in the treatment of ischemic heart diseases. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Computational Model of Aortic Valve Surgical Repair using Grafted Pericardium

    PubMed Central

    Hammer, Peter E.; Chen, Peter C.; del Nido, Pedro J.; Howe, Robert D.

    2012-01-01

    Aortic valve reconstruction using leaflet grafts made from autologous pericardium is an effective surgical treatment for some forms of aortic regurgitation. Despite favorable outcomes in the hands of skilled surgeons, the procedure is underutilized because of the difficulty of sizing grafts to effectively seal with the native leaflets. Difficulty is largely due to the complex geometry and function of the valve and the lower distensibility of the graft material relative to native leaflet tissue. We used a structural finite element model to explore how a pericardial leaflet graft of various sizes interacts with two native leaflets when the valve is closed and loaded. Native leaflets and pericardium are described by anisotropic, hyperelastic constitutive laws, and we model all three leaflets explicitly and resolve leaflet contact in order to simulate repair strategies that are asymmetrical with respect to valve geometry and leaflet properties. We ran simulations with pericardial leaflet grafts of various widths (increase of 0%, 7%, 14%, 21% and 27%) and heights (increase of 0%, 13%, 27% and 40%) relative to the native leaflets. Effectiveness of valve closure was quantified based on the overlap between coapting leaflets. Results showed that graft width and height must both be increased to achieve proper valve closure, and that a graft 21% wider and 27% higher than the native leaflet creates a seal similar to a valve with three normal leaflets. Experimental validation in excised porcine aortas (n=9) corroborates the results of simulations. PMID:22341628

  14. An atlas of bovine gene expression reveals novel distinctive tissue characteristics and evidence for improving genome annotation

    USDA-ARS?s Scientific Manuscript database

    Background A comprehensive transcriptome survey, or gene atlas, provides information essential for a complete understanding of the genomic biology of an organism. We present an atlas of RNA abundance for 92 adult, juvenile and fetal cattle tissues and three cattle cell lines. Results The Bovine Gene...

  15. Stress response of bovine artery and rat brain tissue due to combined translational shear and fixed unconfined compression

    NASA Astrophysics Data System (ADS)

    Leahy, Lauren

    During trauma resulting from impacts and blast waves, sinusoidal waves permeate the brain and cranial arterial tissue, both non-homogeneous biological tissues with high fluid contents. The experimental shear stress response to sinusoidal translational shear deformation at 1 Hz and 25% strain amplitude and either 0% or 33% compression is compared for rat brain tissue and bovine aortic tissue. Both tissues exhibit Mullins effect in shear. Harmonic wavelet decomposition, a novel application to the mechanical response of these tissues, shows significant 1 Hz and 3 Hz components. The 3 Hz component magnitude in brain tissue, which is much larger than in aortic tissue, may correlate to interstitial fluid induced drag forces that decrease on subsequent cycles perhaps because of damage resulting in easier fluid movement. The fluid may cause the quasiperiodic, viscoelastic behavior of brain tissue. The mechanical response differences under impact may cause shear damage between arterial and brain connections.

  16. THE RETENTION OF S35-LABELLED BOVINE SERUM ALBUMIN IN NORMAL AND IMMUNIZED RABBIT LIVER TISSUE

    PubMed Central

    Garvey, Justine S.; Campbell, Dan H.

    1957-01-01

    The S35-label of S35-BSA was detected in the liver tissue of rabbits to the extent of 0.02 per cent (10 µg or ≃ 1014 molecules) of the injected material at 140 days after injection. The rate of loss of antigen at the termination of the experiment was of such an order that significant amounts would be expected to persist for at least several years. Data are reported which extend the retention data previously reported on S35-labelled hemocyanin. They indicate that amounts of the order of 0.05 per cent (25 µg.) of antigen material persist at 330 days after injection. All of the radioactivity of material retained in the liver tissue 6 weeks after injection was immunologically related to the original S35-BSA antigen. Preliminary studies are reported which indicate that the retained antigen is bound to ribonucleic acid. A new method is described for the isolation of p-azophenylsulfonate bovine serum albumin from tissue extracts by means of a Dowex 2 adsorbent. PMID:13416473

  17. Tissue reaction of deproteinized bovine bone matrix grafting in ectopic site: histological study on sheep.

    PubMed

    Grossi, João Ricardo Almeida; Bonacin, Rodrigo; Crivelaro, Viviane Rozeira; Giovanini, Allan Fernando; Zielak, João César; Deliberador, Tatiana Miranda

    2016-12-01

    The aim of this paper was to evaluate through histological analysis of the tissue reaction of deproteinized bovine bone matrix (DBBM) when inserted into the site of intramuscular ectopic sheep. In this study, 16 sheep received 3 groups and these were divided into 2 experimental times: Group 1-sham group, Group 2-particulate autogenous bone and Group 3-DBBM granules. All animals underwent surgical procedures for insertion of materials in an ectopic site (muscles of the lower back and after 3 and 6 months postoperatively, the samples were evaluated by histological analysis. The results indicated that the Sham group showed dense collagen fibers and thin, characterizing fibrosis at 3 and 6 months. In the autograft group there was a significant amount of collagen deposition and decreased inflammation at 6 months postoperatively. Group of DBBM, it was noted the presence of dense connective tissue and surrounding remaining particles was observed the formation of with osteoid characteristic tissue. The DBBM demonstrated biocompatibility, osteoconductivity and small osteogenesis capacity on ectopic site.

  18. Automatic pericardium segmentation and quantification of epicardial fat from computed tomography angiography

    PubMed Central

    Norlén, Alexander; Alvén, Jennifer; Molnar, David; Enqvist, Olof; Norrlund, Rauni Rossi; Brandberg, John; Bergström, Göran; Kahl, Fredrik

    2016-01-01

    Abstract. Recent findings indicate a strong correlation between the risk of future heart disease and the volume of adipose tissue inside of the pericardium. So far, large-scale studies have been hindered by the fact that manual delineation of the pericardium is extremely time-consuming and that existing methods for automatic delineation lack accuracy. An efficient and fully automatic approach to pericardium segmentation and epicardial fat volume (EFV) estimation is presented, based on a variant of multi-atlas segmentation for spatial initialization and a random forest classifier for accurate pericardium detection. Experimental validation on a set of 30 manually delineated computer tomography angiography volumes shows a significant improvement on state-of-the-art in terms of EFV estimation [mean absolute EFV difference: 3.8 ml (4.7%), Pearson correlation: 0.99] with run times suitable for large-scale studies (52 s). Further, the results compare favorably with interobserver variability measured on 10 volumes. PMID:27660804

  19. Biomechanical and structural changes following the decellularization of bovine pericardial tissues for use as a tissue engineering scaffold.

    PubMed

    Pagoulatou, Eirini; Triantaphyllidou, Irene-Eva; Vynios, Demitrios H; Papachristou, Dionysios J; Koletsis, Efstratios; Deligianni, Despina; Mavrilas, Dimosthenis

    2012-06-01

    To achieve natural scaffolds for tissue engineering applications we decellularized bovine pericardial (BP) tissues according to two different protocols: a novel treatment based on Triton(®) X-100 (12 h, 4 °C) (BP1) and a trypsin/EDTA treatment (37 °C, 48 h) (BP2). Results were compared with commercially available acellular xenogeneic biomaterials, Veritas(®) and Collamed(®). Biomechanical characteristics, high (E(h)) and low (E(l)) modulus of elasticity, of the fresh untreated tissue varied with the anatomical direction (apex to base (T) to transverse (L)) (mean ± SDEV): (41.63 ± 14.65-48.12 ± 10.19 MPa and 0.27 ± 0.05-0.30 ± 0.12 MPa respectively). BP1 had no mechanical effect (44.65 ± 19.73-52.67 ± 7.59 MPa and 0.37 ± 0.14-0.37 ± 0.11 MPa, respectively) but BP2 resulted in significant decrease in E(h) and E(l) (20.96 ± 8.17-36.82 ± 3.23 MPa and 0.20 ± 0.06-0.23 ± 0.06 MPa). Hysteresis ratio (h) varied (19-26 % of the loading energy) independently of anatomical direction. Glycosaminoglycans content was unaffected by BP1, while 22 % of chondroitin/dermatan sulphate and 60 % of hyaluronan were removed after BP2 treatment. Endothelial cell adhesion was achieved after 24 h and 3 days cell culture.

  20. Transcriptome analysis of the medulla tissue from cattle in response to bovine spongiform encephalopathy using digital gene expression tag profiling.

    PubMed

    Basu, Urmila; Almeida, Luciane; Olson, N Eric; Meng, Yan; Williams, John L; Moore, Stephen S; Guan, Le Luo

    2011-01-01

    Bovine spongiform encephalopathy (BSE) is a transmissible, fatal neurodegenerative disorder of cattle produced by prions. The use of excessive parallel sequencing for comparison of gene expression in bovine control and infected tissues may help to elucidate the molecular mechanisms associated with this disease. In this study, tag profiling Solexa sequencing was used for transcriptome analysis of bovine brain tissues. Replicate libraries were prepared from mRNA isolated from control and infected (challenged with 100 g of BSE-infected brain) medulla tissues 45 mo after infection. For each library, 5-6 million sequence reads were generated and approximately 67-70% of the reads were mapped against the Bovine Genome database to approximately 13,700-14,120 transcripts (each having at least one read). About 42-47% of the total reads mapped uniquely. Using the GeneSifter software package, 190 differentially expressed (DE) genes were identified (>2.0-fold change, p < .01): 73 upregulated and 117 downregulated. Seventy-nine DE genes had functions described in the Gene Ontology (GO) database and 16 DE genes were involved in 38 different pathways described in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Digital analysis expression by tag profiling may be a powerful approach to comprehensive transcriptome analysis to identify changes associated with disease progression, leading to a better understanding of the underlying mechanism of pathogenesis of BSE.

  1. Computed tomography of the abnormal pericardium

    SciTech Connect

    Silverman, P.M.; Harell, G.S.; Korobkin, M.

    1983-06-01

    Computed tomographic (CT) findings in 18 patients with documented pericardial disease are reported. The pericardium appears as a thin, curvilinear, 1- to 2-mm-thick density best seen anterior to the right ventricular part of the heart. Pericardial abnormalities detected by CT include effusions, thickening, calcification, and cystic and solid masses. Computed tomography is complimentary to echocardiography in its ability to more accurately characterize pericardial effusions, masses, and pericardial thickening.

  2. Quantification of PrPC in bovine peripheral tissues: Analysis in wild-type and PrPC-deficient cattle.

    PubMed

    Kobayashi, Shin-Ichi; Ano, Yasuhisa; Sakudo, Akikazu; Yukawa, Masayoshi; Sigiura, Katsuaki; Manabe, Noboru; Nakayama, Hiroyuki; Onodera, Takashi

    2009-01-01

    Cellular PrP (PrPC) is necessary for bovine spongiform encephalopathy (BSE) infection. The purpose of the present experiment was the quantification of PrPC in peripheral tissues to assess the risk of BSE infection from these tissues. The tissue distribution of PrPC was examined by a sandwich enzyme-linked immunosorbent assay (sELISA) and histochemical analysis. PrPC-deficient cows were used as a negative control. The sELISA revealed that the brain contained the highest PrPC content (10.7 µg/g tissue), while other organs/tissues harbored lower amounts, in decreasing order as follows: longissimus capitis muscle, iliocostalis thoracis muscle, splenius muscle, biceps femoris muscle, triceps brachii muscle, longissimus thoracis muscle, ileum, jejunum, duodenum, colon, cecum, apex linguae, omotransversarius muscle, posterior part of the corpus linguae, anterior part of the corpus linguae and radix linguae (5.2- to 31-fold less PrPC than the brain). In the tissue/organs of PrP-deficient cows, PrPC levels were under the limit of detection. Histochemical analysis showed that PrPC was expressed in nerve cells in intestinal tissues. The presence of PrPC in the bovine tongue, skeletal muscles and intestines raises the possibility of PrPSc accumulation in these tissues, indicating that these organs/tissues may serve as potential sources of BSE infection.

  3. Seamless reconstruction of mitral leaflet and chordae with one piece of pericardium.

    PubMed

    Ito, Toshiaki; Maekawa, Atsuo; Aoki, Masakazu; Hoshino, Satoshi; Hayashi, Yasunari; Sawaki, Sadanari; Yanagisawa, Junji; Tokoro, Masayoshi

    2014-06-01

    Mitral valve repair is challenging when enough pliable mitral leaflets and chordae are not left intact because of extensive infective endocarditis or chronic sclerotic degeneration. For those cases, we developed a simple method to reconstruct defective leaflets and chordae en bloc with a piece of pericardium, and the mid-term results were evaluated. From January 2009 to November 2013, 25 patients with the mean age of 63 (range 20-88) years underwent this operation. The causes of mitral regurgitation were infective endocarditis in 8, sclerotic degeneration in 8, leaflet dehiscence of previous repair in 2, mitral annular calcification in 3, rheumatic in 2 and congenital in 2. After complete debridement of infected or consolidated tissue, we reconstructed defective mitral leaflets and chordae en bloc with a piece of glutaraldehyde-treated autologous pericardium. To substitute posterior leaflet and chordae, the pericardium was trimmed into a narrow pentagonal shape. The pointed end was attached directly to the corresponding papillary muscle, basal side edges to remnant leaflets on both sides, and the base to the annulus. For anterior leaflet, the pericardium was trimmed into a triangular shape if the lesion was confined in the left or right half or into a double-triangle shape if the lesion involved whole anterior leaflet. The summit of triangle was fixed to corresponding papillary muscle, and the base to remnant anterior leaflet, thus reconstructing coaptation zone and chordae seamlessly. There was no hospital death, and mitral regurgitation at discharge was none or trivial in all patients. During 1-59 months (mean 12.7) of complete follow-up, death, infection or hemolysis was not observed. In one patient, mitral regurgitation recurred 8 months postoperatively because the fixation suture of the pericardium to the papillary muscle broke. The valve was re-repaired with re-attaching the leg of the pericardium. Regurgitation was less than moderate in all other patients

  4. Widespread expression of SAA and Hp RNA in bovine tissues after evaluation of suitable reference genes.

    PubMed

    Lecchi, Cristina; Dilda, Francesca; Sartorelli, Paola; Ceciliani, Fabrizio

    2012-01-15

    The serum amyloid A (SAA) and haptoglobin (Hp) are the most prominent acute phase proteins (APPs) in cow. Liver mainly produces APPs, but extra hepatic expression has also been demonstrated in some tissues. The major aim of the present study was to assess the constitutive SAA and Hp mRNA expression by quantitative PCR (qPCR) in a wide panel of 33 bovine tissues, including gastrointestinal tract, respiratory system, urogenital system, mammary gland, hematopoietic system, central nervous system, eye, thyroid and heart. Normalization of gene expression in different samples requires reference genes, which are stably expressed. Therefore, seven reference genes were investigated (ACTB, GAPDH, HMBS, SDHA, YWHAZ, SF3A1, EEF1A2) and three genes, namely SF3A1, HMBS and ACTB, were selected after assessing their stability with geNorm™ and NormFinder(©) softwares. The qPCR analysis confirmed liver as the principal source of SAA and Hp, but also identified both APPs' mRNA in almost all tissues. The highest expression rate of SAA was found in thyroid, followed by pancreas and submandibulary gland. Hp mRNA expression was detected at high concentration in pancreas and submandibulary gland. The present data indicated a widespread expression of SAA and Hp also in non pathological conditions, thus envisaging a possible role as immunomodulatory and protective molecules. To understand where SAA and Hp come from is the prerequisite to their utilization as Acute Phase Reaction biomarkers. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. A survey of malformed aborted bovine fetuses, stillbirths and nonviable neonates for abnormal karyotypes.

    PubMed Central

    Coates, J W; Schmutz, S M; Rousseaux, C G

    1988-01-01

    Postmortem examinations were performed on 30 morphologically abnormal aborted bovine fetuses, stillbirths and nonviable neonates. Fibroblasts from the pericardium were cultured for chromosome analysis. Karyotypes were successfully completed on 18 animals, of which three were trisomic, one was mosaic monosomic and one was chimeric. All aneuploid calves had multisystemic anomalies. Using chromosomal banding techniques, the abnormal karyotypes were determined to be: 61,XY,+27; 61,XX,+21; 61,XY,+?; 59,XY,-?/60,XY; and 60,XX/60,XY. Bacterial contamination or nonviability of tissues prevented the growth of fibroblasts in culture and cytogenetic analysis of the other 12 animals. It was estimated that 2.0% of all late gestation abortuses and stillbirths may have chromosomal abnormalities characterized by aneuploidy. The findings of this study suggest chromosomal abnormalities characterized by aneuploidy are a significant cause of multisystemic anomalies in aborted bovine fetuses and nonviable neonates. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. PMID:3370561

  6. 125 INCOMPLETE COMPENSATORY UP-REGULATION OF X-LINKED GENES IN BOVINE GERMLINE, EARLY EMBRYOS, AND SOMATIC TISSUES.

    PubMed

    Duan, J; Jue, N K; Jiang, Z; O'Neill, R; Wolf, E; Blomberg, L A; Dong, H; Zheng, X; Chen, J; Tian, X

    2016-01-01

    The maintenance of a proper gene dosage is essential in cellular networks. To resolve the dosage imbalance between eutherian females (XX) and male (XY), X chromosome inactivation (XCI) occurs in females, while X-chromosome dosage compensation up-regulates the active X to balance its expression with that of autosome pairs [Ohno's hypothesis; Ohno 1967 Sex Chromosomes and Sex-linked Genes (Springer-Verlag), p. 99]. These phenomena have been well studied in humans and mice, despite many controversies over the existence of such up-regulation. Using RNA sequencing data, we determined X chromosome dosage compensation in the bovine by analysing the global expression profiles of germ cells, embryos, and somatic tissues. Eight bovine RNA-seq data sets were obtained in from the Gene Expression Omnibus, covering bovine immature/mature oocytes (GSE59186 and GSE52415), pre-implantation conceptuses (GSE59186, GSE52415, and GSE56513), extra-embryonic tissues (PRJNA229443), and male/female somatic tissues (GSE74076, GSE63509, PRJEB6377, and GSE65125). The RNAseq data were trimmed and non-uniquely (paralogs included) mapped to the bovine reference genome assembly UMD3.1.1 using Hisat2 (version 2.0.5) aligner. The mRNA level of each gene, estimated by transformed transcripts per kilobase million was quantified by IsoEM (version 1.1.5). These RNA-seq data sets represented 4 chromosome scenarios in cells: XXXX:AAAA (diploid immature oocyte with DNA duplication), XX:AA (haploid mature oocyte with DNA duplication), XX:AA and X:AA (gradual changed X status in bovine pre-implantation conceptuses), and X:AA (extra-embryonic tissues and somatic cells in female with one active X or XY male) were analysed for dosage compensation. A total of 959 X-linked genes and 20,316 autosome genes were used to calculate the relative X to autosomal gene (A) expression (RXE): log2 (X expression) - log2 (A expression). The following dosage determinations were made: RXE values ≥ 0: complete dosage

  7. Bovine muscle 20S proteasome. III: Quantification in tissue crude extracts using ELISA and radial immunodiffusion techniques and practical applications.

    PubMed

    Aubry, L; Sentandreu, M A; Levieux, D; Ouali, A; Dutaud, D

    2006-10-01

    The 20S proteasome is a large complex (700kDa) that exhibits endo- and exo-peptidase activities with wide specificity. In postmortem muscles, several sets of evidence suggest a possible significant contribution of proteasome to meat tenderisation. Hence, an accurate and rapid quantification procedure is needed to attest that new function during the ageing of meat. In the present work, we developed an ELISA test enabling the quantification of nM concentrations of the 20S proteasome. We further tested the radial immunodiffusion (RID) technique described as a more simple method that can quantitatively determine the concentration of an antigen in a complex mixture. The ELISA test allowed us to quantify the 20S protesome in tissue homogenates and fluids with a recovery of 100%, a coefficient of variation lower than 5% and a detection limit of 9ng/ml. Quantification of the 20S proteasome in various bovine tissue by ELISA showed the highest concentration in liver followed by spleen and kidney, with muscles exhibiting the lowest concentrations. In addition, measurement of the proteasome concentration in eight different bovine muscles with various metabolic profiles led to the conclusion that the relationship between muscle metabolic properties and proteasome concentration is rather complex. Nevertheless, heart muscle exhibited the highest proteasome content (331μg/g wet tissue) whereas the lowest values were found for M. Tensor Fascia Latae (213μg/g wet tissue), a fast twitch white muscle, M. Supraspinatus (209μg/g wet tissue), a slow twitch red muscle and M. Pectoralis profondus (203μg/g wet tissue), an intermediate muscle. As compared to other endogenous peptidases, muscle tissue contains relatively high amounts of proteasome. Hence this complex can be quantified using the RID, which allows quantification of protein in the μg range. Plotting the concentration values determined with both methods for all bovine tissues tested gave a straight line with a correlation

  8. Variations in energy metabolism along the pericardium meridian and its relationship with visceral function adjustments during electroacupuncture.

    PubMed

    Zheng, Shu-Xia; Pan, Xiao-Hua; Xu, Jin-Sen; Xiu, Chun-Ying; Dong, Ya-Qin; Zhu, Xiaoxiang

    2014-08-30

    , the decrease in oxygen partial pressure along the pericardium meridian might be a result of strengthened energy metabolism of associated tissue and increased oxygen consumption. The variations in energy metabolism along the pericardium meridian during the course of EA had a close relationship with visceral function adjustments. Chinese Clinical Trial Registry ChiCTRTRC13003193.

  9. Galactosylated bovine serum albumin nanoparticles for parenteral delivery of oridonin: tissue distribution and pharmacokinetic studies.

    PubMed

    Li, Caiyun; Zhang, Dianrui; Guo, Yuanyuan; Guo, Hejian; Li, Tingting; Hao, Leilei; Zheng, Dandan; Liu, Guangpu; Zhang, Qiang

    2014-01-01

    Bovine serum albumin (BSA) nanoparticle is a promising drug carrier system. Oridonin (ORI)-loaded galactosylated BSA nanoparticle (ORI-GB-NP) was prepared for liver targeting delivery of ORI. This work was designed to investigate the in vitro release, in vivo pharmacokinetics and tissue distribution of ORI-GB-NP. ORI-GB-NP was prepared by the desolvation method. The particle size of ORI-GB-NP was 172.0 ± 8.3 nm with narrow size distribution. The in vitro release of ORI-GB-NP exhibited biphasic drug release pattern with an initial burst release and consequently sustained release. Pharmacokinetic analysis displayed that ORI-GB-NP and ORI-loaded BSA nanoparticle (ORI-BSA-NP) could enhance the drug plasma level and prolong the circulation time in contrast with ORI solution. Meanwhile, compared with ORI-BSA-NP, ORI-GB-NP could deliver more ORI to liver and simultaneously reduce the toxicity of ORI to heart, lung and kidney. In conclusion, ORI-GB-NP could be a promising drug delivery system for liver cancer therapy.

  10. Exposure of bovine dermal tissue to ultraviolet light under the Antarctic ozone hole

    NASA Astrophysics Data System (ADS)

    Takahashi, Tetsuya; Ogura, Takayuki; Tanaka, Keisuke; Hattori, Shunji; Kudoh, Sakae; Imura, Satoshi

    2016-12-01

    Bovine dermis was exposed outdoors in the Antarctic in 2013 to study the skin damage caused by short-wavelength ultraviolet light under the ozone hole. Collagen was extracted from the exposed dermis with pepsin. The amount of solubilized collagen in the exposed dermis was only 20%-40% of that in dermis shielded from ultraviolet light. The dermis was most difficult to extract when exposed in summer, and then when exposed in spring. Differential scanning calorimetry was used to determine the melting endothermic behavior of the dermal tissue. The peak temperature was highest for the dermis exposed in summer. The exposed dermis was degraded with cyanogen bromide to determine whether cross-linking had occurred. Cross-linked peptides were detected in the dermis exposed in summer or spring, but the dermis exposed in autumn did not differ markedly from the light-shielded dermis. These data show that cross-linkages were readily formed in the collagen molecule chains in dermis exposed to ultraviolet light in summer, when solar elevation is highest and the period of sunshine is longest. A comparison of the dermis exposed in spring and that exposed in autumn showed that cross-linkages were formed more readily by ultraviolet light in spring, when the ozone hole occurred.

  11. Evaluation of the thermal property of bovine intramuscular adipose tissue using differential scanning calorimetry.

    PubMed

    Kimura, Noriyuki; Nishimura, Nana; Iwama, Nagako; Aihara, Yoshito; Ogawa, Yasuki; Miyaguchi, Yuji

    2017-10-01

    The thermal property of bovine intramuscular adipose tissue (IAT) was evaluated using differential scanning calorimetry (DSC) and compared with the melting point temperature (MP) of the fat extract of IAT, which was measured using the slip point method. The beef samples were classified according to the beef marbling score (BMS). Beef with a high BMS contained less protein than that with middle or low BMS. Beef with a high BMS contained significantly more fat than that with a low BMS (P < 0.05). The endothermic point temperature (EP) of IAT, measured by DSC, was significantly higher than the MP of IAT fat (P < 0.05). The EP showed no significant difference among the three marbling grade groups. Although the MP was correlated with the monounsaturated fatty acids (MUFA) content of IAT (R(2)  = 0.505), there was no correlation between the EP and the MUFA (R(2)  = 0.040). However, the EP of IAT treated with collagenase was relatively highly correlated with the MP (R(2)  = 0.655). Thus, these results suggested that DSC analysis would give us the practical thermal information regarding the melt-in the-mouth of beef such as the gelatinization of collagen, along with the melting of fat in IAT. © 2017 The Authors. Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.

  12. On-line MSPD-SPE-HPLC/FLD analysis of polycyclic aromatic hydrocarbons in bovine tissues.

    PubMed

    Gutiérrez-Valencia, Tania M; García de Llasera, Martha P

    2017-05-15

    A fast method was optimized and validated for simultaneous trace determination of four polycyclic aromatic hydrocarbons: benzo[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene and benzo[a]pyrene in bovine tissues. The determination was performed by matrix solid-phase dispersion (MSPD) coupled on-line to solid phase extraction (SPE) and high performance liquid chromatography (HPLC) with fluorescence detection (FLD). The sample was dispersed on C18 silica sorbent and then the on-line MSPD-SPE-HPLC/FLD method was applied. Several parameters were optimized: cleaning and elution sequences applied to the MSPD cartridge, the flow rate and dilution of extract used for SPE loading. The on-line method was validated over a concentration range of 0.1-0.6ngg(-1) obtaining good linearity (r⩾0.998) and precision (RSD)⩽10%. Recovery ranged from 96 to 99% and the limits of detection were 0.012ngg(-1). This methodology was applied to liver samples from unhealthy animals. The results demonstrate that MSDP-SPE-HPLC/FLD method provides reliable, sensitive, accurate and fast data to the food control.

  13. Detection of bovine central nervous system tissues in rendered animal by-products by one-step real-time reverse transcription PCR assay.

    PubMed

    Andrievskaia, Olga; Tangorra, Erin

    2014-12-01

    Contamination of rendered animal byproducts with central nervous system tissues (CNST) from animals with bovine spongiform encephalopathy is considered one of the vehicles of disease transmission. Removal from the animal feed chain of CNST originated from cattle of a specified age category, species-labeling of rendered meat products, and testing of rendered products for bovine CNST are tasks associated with the epidemiological control of bovine spongiform encephalopathy. A single-step TaqMan real-time reverse transcriptase (RRT) PCR assay was developed and evaluated for specific detection of bovine glial fibrillary acidic protein (GFAP) mRNA, a biomarker of bovine CNST, in rendered animal by-products. An internal amplification control, mammalian b -actin mRNA, was coamplified in the duplex RRT-PCR assay to monitor amplification efficiency, normalize amplification signals, and avoid false-negative results. The functionality of the GFAP mRNA RRT-PCR was assessed through analysis of laboratory-generated binary mixtures of bovine central nervous system (CNS) and muscle tissues treated under various thermal settings imitating industrial conditions. The assay was able to detect as low as 0.05 % (wt/wt) bovine brain tissue in binary mixtures heat treated at 110 to 130°C for 20 to 60 min. Further evaluation of the GFAP mRNA RRT-PCR assay involved samples of industrial rendered products of various species origin and composition obtained from commercial sources and rendering plants. Low amounts of bovine GFAP mRNA were detected in several bovine-rendered products, which was in agreement with declared species composition. An accurate estimation of CNS tissue content in industrial-rendered products was complicated due to a wide range of temperature and time settings in rendering protocols. Nevertheless, the GFAP mRNA RRT-PCR assay may be considered for bovine CNS tissue detection in rendered products in combination with other available tools (for example, animal age

  14. Short communication: Endoplasmic reticulum stress gene network expression in bovine mammary tissue during the lactation cycle.

    PubMed

    Invernizzi, G; Naeem, A; Loor, J J

    2012-05-01

    The endoplasmic reticulum (ER) has a crucial role in cellular metabolism. Recent studies in nonruminants discovered that components of the ER stress pathway, induced during the unfolded protein response, play critical roles in regulating lipogenesis. The bovine mammary gland faces extreme metabolic stress at the onset of lactation due primarily to the increase in flux through pathways associated with milk fat and protein synthesis. Our objective was to study, via quantitative real-time PCR, the expression of the ER stress pathway components P58IPK, PERK, XBP1, ATF4, ATF3, ATF6, CHOP, MBTPS1, GRP94, and BiP in mammary tissue (n=7 cows × 5 time points) collected at -15, 1, 15, 60, and 240 d relative to parturition. Expression of P58IPK and ATF4 increased to a peak at d 60, followed by a decrease by d 240 postpartum. Despite the decrease in expression by 240 d, P58IPK remained higher than prepartal levels (d -15). Expression patterns of ATF3 and CHOP were similar and peaked at d 15, followed by a decrease through d 240, at which point CHOP expression was still greater than prepartal levels. The sharp increase in milk production postpartum (d 15) as well as apoptosis during late lactation (240 d) may have induced a pseudo unfolded protein response state. This is supported by the similar expression patterns of P58IPK and PERK. In the context of lactation, however, transcriptional changes in the ER stress pathway at different stages of the lactation cycle are a normal aspect of the tissue's adaptation to the changing physiological state. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Agouti revisited: transcript quantification of the ASIP gene in bovine tissues related to protein expression and localization.

    PubMed

    Albrecht, Elke; Komolka, Katrin; Kuzinski, Judith; Maak, Steffen

    2012-01-01

    Beside its role in melanogenesis, the agouti signaling protein (ASIP) has been related to obesity. The potentially crucial role in adipocyte development makes it a tempting candidate for economic relevant, fat related traits in farm animals. The objective of our study was to characterize the mRNA expression of different ASIP transcripts and of putative targets in different bovine tissues, as well as to study consequences on protein abundance and localization. ASIP mRNA abundance was determined by RT-qPCR in adipose and further tissues of cattle representing different breeds and crosses. ASIP mRNA was up-regulated more than 9-fold in intramuscular fat of Japanese Black cattle compared to Holstein (p<0.001). Further analyses revealed that a transposon-derived transcript was solely responsible for the increased ASIP mRNA abundance. This transcript was observed in single individuals of different breeds indicating a wide spread occurrence of this insertion at the ASIP locus in cattle. The protein was detected in different adipose tissues, skin, lung and liver, but not in skeletal muscle by Western blot with a bovine-specific ASIP antibody. However, the protein abundance was not related to the observed ASIP mRNA over-expression. Immuno-histochemical analyses revealed a putative nuclear localization of ASIP additionally to the expected cytosolic signal in different cell types. The expression of melanocortin receptors (MCR) 1 to 5 as potential targets for ASIP was analyzed by RT-PCR in subcutaneous fat. Only MC1R and MC4R were detected indicating a similar receptor expression like in human adipose tissue. Our results provide evidence for a widespread expression of ASIP in bovine tissues at mRNA and, for the first time, at protein level. ASIP protein is detectable in adipocytes as well as in further cells of adipose tissue. We generated a basis for a more detailed investigation of ASIP function in peripheral tissues of various mammalian species.

  16. Sugar binding to purified fractions from bovine taste buds and epithelial tissue. Relationships to bioactivity.

    PubMed

    Lum, C K; Henkin, R I

    1976-02-24

    Binding of various sugars was compared in purified subfractions of taste buds isolated from bovine circumvallate papillae and of non-taste bud-bearing epithelium isolated from tissue surrounding these papillae. Binding of 14C-labeled sugars was greater in purified subfractions obtained from taste bud than from non-taste bud-bearing tissue and was, in general, greater in those taste bud subfractions in which a greater membrane purification was achieved. Binding specificity of the 14C-labeled sugars sucrose, fructose, glucose and of 14C-labeled cyclamate and saccharine was measured by competition of each 14C-labeled sugar or synthetic sweetener with its unlabeled homologous sugar in P4(B) taste bud subfractions; this binding, as shown for sucrose, was reversible and temperature dependent. Essentially no competition of the 14C-lageled sugars sucrose, fructose, glucose or 14C-labeled cyclamate and saccharine by their respective unlabeled homologues occurred in epithelial tissue P4(B) subfractions; this binding was not reversible. Binding specificity was further observed by the competition of 14C-labeled sucrose, fructose and glucose with each unlabeled sugar for binding sites on P4(B) taste bud subfractions; unlabeled sucrose was more effective in competing with each 14C-labeled surgar than was unlabeled fructose or glucose. The relatively non-sweet sugar lactose did not compete with 14C-labeled lactose in P4(B) subfractions from either taste bud or non-taste bud-bearing epithelial tissue. Binding of 14C-labeled sucrose in purified P4(B) bud subfractions was inhibited by increased concentrations of unlabeled sucrose, phospholipase C, neuraminidase, EDTA, NaCl and urea. Dissociation constants for sugar or synthetic sweetener binding were low (approx. 10(-3) M) but in a rank order (sucrose greater than fructose greater than glucose greater than saccharine) consistent with preference and electrophysiological responses in cow. The cow is behaviorally indifferent to

  17. TRIENNIAL GROWTH AND DEVELOPMENT SYMPOSIUM: Factors influencing bovine intramuscular adipose tissue development and cellularity.

    PubMed

    Albrecht, E; Schering, L; Liu, Y; Komolka, K; Kühn, C; Wimmers, K; Gotoh, T; Maak, S

    2017-05-01

    Appearance, distribution, and amount of intramuscular fat (IMF), often referred to as marbling, are highly variable and depend on environmental and genetic factors. On the molecular level, the concerted action of several drivers, including hormones, receptors, transcription factors, etc., determines where clusters of adipocytes arise. Therefore, the aim of future studies remains to identify such factors as biological markers of IMF to increase the ability to identify animals that deposit IMF early in age to increase efficiency of high-quality meat production. In an attempt to unravel the cellular development of marbling, we investigated the abundance of markers for adipogenic differentiation during fattening of cattle and the transcriptome of muscle and dissected IMF. Markers of different stages of adipogenic differentiation are well known from cell culture experiments. They are usually transiently expressed, such as delta-like homolog 1 (DLK1) that is abundant in preadipocytes and absent during differentiation to mature adipocytes. It is even a greater challenge to detect those markers in live animals. Within skeletal muscles, hyperplasia and hypertrophy of adipocytes can be observed throughout life. Therefore, development of marbling requires, on the cellular level, recruitment, proliferation, and differentiation of adipogenic cells to store excess energy in the form of lipids in new cells. In a recent study, we investigated the localization and abundance of early markers of adipogenic differentiation, such as DLK1, in bovine muscle tissue. An inverse relationship between IMF content and number of DLK1-positive cells in bovine muscle was demonstrated. Considering the cellular environment of differentiating adipocytes in muscle and the secretory action of adipocytes and myocytes, it becomes obvious that cross talk between cells via adipokines and myokines may be important for IMF development. Secreted proteins can act on other cells, inhibiting or stimulating

  18. Antioxidant adaptive responses of extraembryonic tissues from cloned and non-cloned bovine conceptuses to oxidative stress during early pregnancy.

    PubMed

    Al-Gubory, Kaïs H; Garrel, Catherine; Delatouche, Laurent; Heyman, Yvan; Chavatte-Palmer, Pascale

    2010-07-01

    Placental oxidative stress has been suggested as a key factor in early pregnancy failure. Abnormal placental development limits success in pregnancies obtained by somatic cell nuclear transfer (SCNT). Malondialdehyde (MDA) content, an index of oxidative stress, and superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities were determined in bovine extraembryonic tissues of SCNT or artificial insemination (AI) conceptuses. Chorionic tissues of SCNT and AI conceptuses show no difference in MDA content at day 32 of pregnancy. MDA content in chorionic tissues of SCNT and AI conceptuses decreased from day 32 to 62 of pregnancy. MDA content was lower in chorionic tissues of SCNT conceptuses than that in chorionic tissues of AI conceptuses at day 62 of pregnancy. SOD1, SOD2 and GPX activities in chorionic tissues of SCNT conceptuses were not different from those in chorionic tissues of AI conceptuses at both gestational ages. CAT activity in chorionic tissues of SCNT conceptuses was lower at day 32, and it was higher at day 62 of pregnancy than that in chorionic tissues of AI conceptuses. CAT and GPX activities increased in chorionic tissues of SCNT conceptuses with gestational age. SOD1 activity decreased while that of SOD2 and GPX increased in chorionic tissues of AI conceptuses with gestational age. At day 62 of pregnancy, MDA content and enzyme activities in cotyledonary tissues were not different between AI and SCNT conceptuses. Different antioxidant mechanisms may operate within the chorion of AI and SCNT conceptuses. Further experiments are required to elucidate this point.

  19. Tissue expression analysis, cloning and characterization of the 5'-regulatory region of the bovine FABP3 gene.

    PubMed

    Li, Anning; Wu, Lijuan; Wang, Xiaoyu; Xin, Yaping; Zan, Linsen

    2016-09-01

    Fatty acid binding protein 3 (FABP3) is a member of the FABP family which bind fatty acids and have an important role in fatty acid metabolism. A large number of studies have shown that the genetic polymorphisms of FABP3 are positively correlated with intramuscular fat (IMF) content in domestic animals, however, the function and transcriptional characteristics of FABP3 in cattle remain unclear. Real-time PCR analysis revealed that bovine FABP3 was highly expressed in cardiac tissue. The 5'-regulatory region of bovine FABP3 was cloned and its transcription initiation sites were identified. Sequence analysis showed that many transcriptional factor binding sites including TATA-box and CCAAT-box were present on the 5'-flanking region of bovine FABP3, and four CpG islands were found on nucleotides from -891 to +118. Seven serial deletion constructs of the 5'-regulatory region evaluated in dual-luciferase reporter assay indicated that its core promoter was 384 base pairs upstream from the transcription initiation site. The transcriptional factor binding sites RXRα, KLF15, CREB and Sp1 were conserved in the core promoter of cattle, sheep, pigs and dogs. These results provide further understanding of the function and regulation mechanism of bovine FABP3.

  20. An atlas of bovine gene expression reveals novel distinctive tissue characteristics and evidence for improving genome annotation

    PubMed Central

    2010-01-01

    Background A comprehensive transcriptome survey, or gene atlas, provides information essential for a complete understanding of the genomic biology of an organism. We present an atlas of RNA abundance for 92 adult, juvenile and fetal cattle tissues and three cattle cell lines. Results The Bovine Gene Atlas was generated from 7.2 million unique digital gene expression tag sequences (300.2 million total raw tag sequences), from which 1.59 million unique tag sequences were identified that mapped to the draft bovine genome accounting for 85% of the total raw tag abundance. Filtering these tags yielded 87,764 unique tag sequences that unambiguously mapped to 16,517 annotated protein-coding loci in the draft genome accounting for 45% of the total raw tag abundance. Clustering of tissues based on tag abundance profiles generally confirmed ontology classification based on anatomy. There were 5,429 constitutively expressed loci and 3,445 constitutively expressed unique tag sequences mapping outside annotated gene boundaries that represent a resource for enhancing current gene models. Physical measures such as inferred transcript length or antisense tag abundance identified tissues with atypical transcriptional tag profiles. We report for the first time the tissue-specific variation in the proportion of mitochondrial transcriptional tag abundance. Conclusions The Bovine Gene Atlas is the deepest and broadest transcriptome survey of any livestock genome to date. Commonalities and variation in sense and antisense transcript tag profiles identified in different tissues facilitate the examination of the relationship between gene expression, tissue, and gene function. PMID:20961407

  1. Tissue concentration of intrauterine and intravenously administered amoxicillin in the isolated hemoperfused bovine uterus model and in healthy cows.

    PubMed

    Braun, Michael; Zerbe, Holm; Kietzmann, Manfred

    2009-01-01

    The tissue distribution of amoxicillin in the bovine uterus in dependence of the administration route was examined using the in vitro model of the isolated hemoperfused bovine uterus. Four hours after intrauterine instillation or onset of perfusion with amoxicillin ("systemic" treatment), respectively, samples of the uterine tissue were taken from different locations of the organ. The tissue concentration of amoxicillin in the single layers of the uterine wall was determined via high performance liquid chromatography (HPLC) analysis. To compare the results with data achieved in vivo, healthy cows were treated post partum intrauterinely or systemically, respectively, and the concentration of amoxicillin was determined in endometrial biopsies. In vitro intrauterine instillation resulted in significantly higher levels of amoxicillin in the inner layers of the uterine wall compared to systemic treatment, whereas the drug is distributed more homogenously through all layers in the systemically treated organs. In the in vivo experiments the tissue concentration of amoxicillin is significantly lower after intrauterine instillation compared to the in vitro data. In contrast, the endometrial concentrations in the uteri perfused with amoxicillin in vitro are comparable to those measured in endometrium of systemically treated cows. The MIC for gram positive and negative bacteria was exceeded in the uterine wall of all organs in vitro and in vivo after intrauterine treatment. In systemic treated cows the MIC for gram negative bacteria was exceeded in two of three cases, but never in the hemoperfused uteri. The data indicate that combined systemic and intrauterine treatment results in the best distribution of sufficient antibiotic concentrations in the uterine tissue. The isolated hemoperfused bovine uterus is shown to be suitable as an in vitro model for pharmacokinetic examinations, especially with regard to detailed information on the tissue distribution of drugs.

  2. Quantification of the edge effect in calcified bioprosthetic tissues.

    PubMed

    Wika, K E; Utoh, J; Brown, J; Harasaki, H

    1993-10-01

    In bioprosthetic tissue samples that had been implanted in the subcutaneous space of rats, and recurring pattern of calcification was observed. In this pattern, which we call the edge effect, the interior of the tissue is calcified and is surrounded and separated from the subcutaneous fluid by a zone that is free from calcification. The edge effect has been qualitatively described in the literature for subcutaneous implants and for valve leaflets, and it may be related to the mechanism of calcification for these materials. The thickness of the calcification free outer layer was quantified for glutaraldehyde treated bovine pericardium, glycerol treated bovine pericardium, glutaraldehyde treated human dura mater, and glycerol treated human dura mater. The edge effect values were found to be unique and consistent for each material type, and they were inversely related to the shrinkage temperatures and the calcium contents of the materials. It was determined that the chemical treatment was more important than the tissue type in determining the edge effect value.

  3. Microwave ablation of ex vivo bovine tissues using a dual slot antenna with a floating metallic sleeve.

    PubMed

    Ibitoye, Ayo Zaccheaus; Nwoye, Ephraim Okeke; Aweda, Adebayo Moses; Oremosu, Ademola A; Anunobi, Chidozie Charles; Akanmu, Nurudeen Olanrewaju

    2016-12-01

    To study the efficiency of a dual slot antenna with a floating metallic sleeve on the ablation of different ex vivo bovine tissues. COMSOL Multiphysics® version 4.4 (Stockholm, Sweden), which is based on finite element methods (FEM), was used to design and simulate monopole and dual slot with sleeve antennas. Power, specific absorption rate (SAR), temperature and necrosis distributions in the selected tissues were determined using these antennas. Monopole and dual slot with sleeve antennas were designed, simulated, constructed and applied in this study based on a semi-rigid coaxial cable. Ex vivo experiments were performed on liver, lung, muscle and heart of bovine obtained from a public animal slaughter house. The microwave energy was delivered using a 2.45 GHz solid-state microwave generator at 40 W for 3, 5 and 10 min. Aspect ratio, ablation length and ablation diameter were also determined on ablated tissues and compared with simulated results. Student's t-test was used to compare the statistically significant difference between the performance of the two antennas. The dual slot antenna with sleeve produces localised microwave energy better than the monopole antenna in all ablated tissues using simulation and experimental validation methods. There were significant differences in ablation diameter and aspect ratio between the sleeve antenna and monopole antenna. Additionally, there were no significant differences between the simulation and experimental results. This study demonstrated that the dual slot antenna with sleeve produced larger ablation zones and higher sphericity index in ex vivo bovine tissues with minimal backward heating when compared with the monopole antenna.

  4. Detection of Mycobacterium bovis in bovine and bubaline tissues using nested-PCR for TbD1.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Kláudia S G; Ramos, Carlos Alberto N; Filho, Antonio Francisco S; Vidal, Carlos Eugênio S; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F; Júnior, Antônio A F; Silva, Marcio R; Neto, José Diomedes B; Cerqueira, Valíria D; Zumárraga, Martín J; Araújo, Flábio R

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  5. Detection of Mycobacterium bovis in Bovine and Bubaline Tissues Using Nested-PCR for TbD1

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A. R.; Jorge, Kláudia S. G.; Ramos, Carlos Alberto N.; Filho, Antonio Francisco S.; Vidal, Carlos Eugênio S.; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F.; Júnior, Antônio A. F.; Silva, Marcio R.; Neto, José Diomedes B.; Cerqueira, Valíria D.; Zumárraga, Martín J.; Araújo, Flábio R.

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:24618787

  6. Effects of Rhodomyrtus tomentosa Leaf Extract on Staphylococcal Adhesion and Invasion in Bovine Udder Epidermal Tissue Model.

    PubMed

    Mordmuang, Auemphon; Shankar, Shiv; Chethanond, Usa; Voravuthikunchai, Supayang Piyawan

    2015-10-15

    Bovine mastitis is one of the most important infectious diseases in dairy herds, and staphylococci are the most important etiologic agents of this disease. Antibiotics and chemical agents used in livestock for prevention and cure of the disease can accumulate in milk and give rise to food safety concerns. Rhodomyrtus tomentosa leaf extract was studied as an alternative approach to reduce the bacterial infections. The ethanolic extract of this plant demonstrated antibacterial activity with minimum inhibitory concentration (MIC) values as low as 16-64 μg/mL against staphylococcal isolates. In addition, the extract had an effect on the bacterial cell surface properties by increasing its hydrophobicity in a concentration dependent manner. To further extend the antibacterial efficacy, silver nanoparticles synthesized with the extract, a pure rhodomyrtone, and liposomal encapsulated rhodomyrtone were applied and their inhibitory effects on bacterial adhesion and invasion were determined by ex vivo study in a bovine udder epidermal tissue model. These agents exerted remarkable antibacterial activity against staphylococci and decreased the adhesion of the bacterial cells to the tissues. These results supported that R. tomentosa ethanolic extract could be applied as an alternative agent for bovine udder care in dairy farms.

  7. Effects of Rhodomyrtus tomentosa Leaf Extract on Staphylococcal Adhesion and Invasion in Bovine Udder Epidermal Tissue Model

    PubMed Central

    Mordmuang, Auemphon; Shankar, Shiv; Chethanond, Usa; Voravuthikunchai, Supayang Piyawan

    2015-01-01

    Bovine mastitis is one of the most important infectious diseases in dairy herds, and staphylococci are the most important etiologic agents of this disease. Antibiotics and chemical agents used in livestock for prevention and cure of the disease can accumulate in milk and give rise to food safety concerns. Rhodomyrtus tomentosa leaf extract was studied as an alternative approach to reduce the bacterial infections. The ethanolic extract of this plant demonstrated antibacterial activity with minimum inhibitory concentration (MIC) values as low as 16–64 μg/mL against staphylococcal isolates. In addition, the extract had an effect on the bacterial cell surface properties by increasing its hydrophobicity in a concentration dependent manner. To further extend the antibacterial efficacy, silver nanoparticles synthesized with the extract, a pure rhodomyrtone, and liposomal encapsulated rhodomyrtone were applied and their inhibitory effects on bacterial adhesion and invasion were determined by ex vivo study in a bovine udder epidermal tissue model. These agents exerted remarkable antibacterial activity against staphylococci and decreased the adhesion of the bacterial cells to the tissues. These results supported that R. tomentosa ethanolic extract could be applied as an alternative agent for bovine udder care in dairy farms. PMID:26501314

  8. Evidence of presence of Mycobacterium tuberculosis in bovine tissue samples by multiplex PCR: possible relevance to reverse zoonosis.

    PubMed

    Mittal, M; Chakravarti, S; Sharma, V; Sanjeeth, B S; Churamani, C P; Kanwar, N S

    2014-04-01

    Bovine tuberculosis, caused by Mycobacterium bovis, remains one of the most important zoonotic health concerns worldwide. The transmission of Mycobacterium tuberculosis from humans to animals also occurs especially in countries where there is close interaction of humans with the animals. In the present study, thirty bovine lung tissue autopsy samples from an organized dairy farm located in North India were screened for the presence of Mycobacterium tuberculosis complex by smear microscopy, histopathological findings and PCR. Differential diagnosis of M. tuberculosis and M. bovis was made based on the deletion of mce-3 operon in M. bovis. The present study found eight of these samples positive for M. tuberculosis by multiplex PCR. Sequencing was performed on two PCR-positive representative samples and on annotation, and BLAST analysis confirmed the presence of gene fragment specific to Mycobacterium tuberculosis. The presence of M. tuberculosis in all the positive samples raises the possibility of human-to-cattle transmission and possible adaptation of this organism in bovine tissues. This study accentuates the importance of screening and differential diagnosis of Mycobacterium tuberculosis complex in humans and livestock for adopting effective TB control and eradication programmes. © 2014 Blackwell Verlag GmbH.

  9. Keratolinin: the soluble substrate of epidermal transglutaminase from human and bovine tissue.

    PubMed Central

    Zettergren, J G; Peterson, L L; Wuepper, K D

    1984-01-01

    Substrates of human and bovine epidermal transglutaminase (glutaminyl-peptide gamma-glutamyltransferase, R-glutaminyl-peptide:amine-gamma-glutamyltransferase, EC 2.3.2.13) were isolated and purified by ion exchange chromatography and preparative zone electrophoresis. These substrates of Mr 36,000, which we propose to call keratolinin, incorporated dansylcadaverine and were precipitated by antibody. Keratolinin is ultimately polymerized on the inner leaflet of the keratinocyte membrane to form the cornified envelope. Each Mr 36,000 substrate was dissociated by chaotropic agents or detergents into noncovalent subunits; the Mr of these subunits was 6,000-6,200 on electrophoresis in 15% acrylamide/1% NaDodSO4/6 M urea gels. Isoelectric focusing of human or bovine keratolinin revealed two moieties separated by 0.3-0.4 pH unit (human, 5.4/5.0; bovine, 6.3/6.0). The two proteins were readily resolved by chromatofocusing and each isoelectric moiety of bovine keratolinin incorporated dansylcadaverine by epidermal transglutaminase and calcium and reacted with identity to antiserum to soluble Mr 36,000 keratolinin. Antiserum to human keratolinin failed to crossreact with its bovine counterpart. Antiserum to involucrin did not crossreact with either keratolinin or epidermis by immunodiffusion. Human and bovine epidermal keratolinins are biochemically similar but immunochemically distinct proteins from the epidermis. Involucrin appears only in significant quantities in cell culture. Images PMID:6141559

  10. Primary malignancies of the heart and pericardium.

    PubMed

    Burazor, Ivana; Aviel-Ronen, Sarit; Imazio, Massimo; Markel, Gal; Grossman, Yoni; Yosepovich, Ady; Adler, Yehuda

    2014-09-01

    Primary malignancies of the heart and pericardium are rare. All the available data come from autopsy studies, case reports, and, in recent years, from large, specialized, single-center studies. Nevertheless, if primary malignancy is present, it may have a devastating implication for patients. Malignancies may affect heart function, also causing left-sided or right-sided heart failure. In addition, they can be responsible for embolic events or arrhythmias. Today, with the widespread use of noninvasive imaging modalities, heart tumors become evident, even as an incidental finding. A multimodality imaging approach is usually required to establish the final diagnosis. Despite the increased awareness and improved diagnostic techniques, clinical manifestations of primary malignancy of the heart and pericardium are so variable that their occurrence may still come as a surprise during surgery or autopsy. No randomized clinical trials have been carried out to determine the optimal therapy for these primary malignancies. Surgery is performed for small tumors. Chemotherapy and radiation therapy can be of help. Partial resection of large neoplasms is performed to relieve mechanical effects, such as cardiac compression or hemodynamic obstruction. Most patients present with marginally resectable or technically nonresectable disease at the time of diagnosis. It seems that orthotopic cardiac transplantation with subsequent immunosuppressive therapy may represent an option for very carefully selected patients. Early diagnosis and radical exeresis are of great importance for long-term survival of a primary cardiac malignancy. This can rarely be accomplished, and overall results are very disappointing. © 2014 Wiley Periodicals, Inc.

  11. Hyphenated liquid chromatographic method for the determination of colistin residues in bovine tissues.

    PubMed

    Decolin, D; Leroy, P; Nicolas, A; Archimbault, P

    1997-12-01

    A selective and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the measurement of colistin residues in milk and in four bovine tissues (i.e., muscle, liver, kidney, and fat). The sample treatment consists of protein precipitation using 10% (w/v) trichloroacetic acid, solid-phase purification on C18 cartridges, and precolumn derivatization of colistin with ortho-phthalaldehyde and 2-mercaptoethanol in borate buffer (pH 10.5). This latter step is performed automatically, and the resulting reaction mixture is injected into a switching HPLC system including a precolumn and an analytical column packed with end-capped LiChrospher RP18 (5 microns). Washing the precolumn and final elution onto the analytical column are conducted using acetonitrile-0.01M phosphate buffer (pH 7.0) mixtures with respective proportions of 65:35 and 68:32 (v/v). Detection is carried out by spectrofluorometry (excitation wavelength, 340 nm; emission wavelength, 440 nm). The retention times of the derivatives corresponding to the two main components of colistin (i.e., polymyxins E2 and E1) are approximately 14 and 18 min, respectively. The structural study of the derivatives corresponding to polymyxins E1 and E2 is carried out by HPLC coupled with electrospray mass spectrometry; data obtained confirms that the derivatization process occurs with the five amino groups of the analytes. Selectivity is obtained in the HPLC system versus other coadministered anti-infective drugs (beta-lactams, aminoglycosides, tetracyclines, and sulphonamides) and endogenous compounds. Quantitation is performed using the sum of the peak areas of polymyxin E1 and polymyxin E2 derivatives. Testing linearity affords correlation coefficients greater than 0.990 for calibration curves in the range of 10-500 microL/L for milk, 50-1000 micrograms/kg for muscle and fat, and 100-1000 micrograms/kg for kidney and liver. Relative standard deviation values are less than 10% at a concentration

  12. Bovine pulp tissue dissolution ability of HealOzone®, Aquatine Alpha Electrolyte® and sodium hypochlorite.

    PubMed

    Rossi-Fedele, Giampiero; Steier, Liviu; Dogramaci, Esma Jane; Canullo, Luigi; Steier, Gabriela; de Figueiredo, Jose Antonio Poli

    2013-08-01

    The aim of this study was to evaluate the bovine pulp tissue dissolution ability of HealOzone, Aquatine Alpha Electrolyte® and 0.5% sodium hypochlorite, used alone or in combination. Thirty bovine pulp fragments were weighed, divided into six groups and placed individually in Eppendorf tubes containing the tested solution until total dissolution occurred. The groups were: G1: saline (negative control), G2: Aquatine Alpha Electrolyte®, G3: 0.5% NaOCl (positive control), G4: Saline + HealOzone, G5: 0.5% NaOCl + HealOzone, G6: Aquatine Alpha Electrolyte® + HealOzone. HealOzone was activated for 2 min with a #6 cup covering the test tube opening on a fixed platform. Two blinded observers using 2× loupes magnification assessed the samples continuously for the first 2 h, and then every hour for the next 8 h. Dissolution speed was calculated by dividing pulp weight by dissolution time (mg min(-1) ). G3 (NaOCl) and G5 (NaOCl + HealOzone) dissolved the pulp tissue completely. The mean dissolution speed for G3 was 0.396 mg min(-1) (SD 0.032) and for G5 was 0.775 mg min(-1) (SD 0.2). Student's t-test showed that G5 dissolved bovine pulp tissue faster than G3 (P = 0.01). Only groups containing sodium hypochlorite dissolved pulp tissue, whilst HealOzone enhanced speed of dissolution.

  13. Characterisation and expression profile of the bovine cathelicidin gene repertoire in mammary tissue

    PubMed Central

    2014-01-01

    Background Cathelicidins comprise a major group of host-defence peptides. Conserved across a wide range of species, they have several functions related to host defence. Only one cathelicidin has been found in humans but several cathelicidin genes occur in the bovine genome. We propose that these molecules may have a protective role against mastitis. The aim of this study was to characterise the cathelicidin gene-cluster in the bovine genome and to identify sites of expression in the bovine mammary gland. Results Bioinformatic analysis of the bovine genome (BosTau7) revealed seven protein-coding cathelicidin genes, CATHL1-7, including two identical copies of CATHL4, as well as three additional putative cathelicidin genes, all clustered on the long arm of chromosome 22. Six of the seven protein-coding genes were expressed in leukocytes extracted from milk of high somatic cell count (SCC) cows. CATHL5 was expressed across several sites in the mammary gland, but did not increase in response to Staphylococcus aureus infection. Conclusions Here, we characterise the bovine cathelicidin gene cluster and reconcile inconsistencies in the datasets of previous studies. Constitutive cathelicidin expression in the mammary gland suggests a possible role for these host defence peptides its protection. PMID:24524771

  14. Isolated Partial Congenital Absence of the Pericardium: A Familial Presentation.

    PubMed

    Palau, Patricia; Domínguez, Eloy; García-González, Pilar; Gallego, Jorge; Bosch, María José; Sieso, Eduardo

    2016-08-01

    Congenital defects of the pericardium are uncommon heart abnormalities. Most of the patients are asymptomatic and are usually diagnosed incidentally. Complications are more common in partial absence than in complete absence of the pericardium; thus, this congenital defect should be identified because of the associated risk of sudden death. We report the first mention in the literature, to our knowledge, of a 3-generation familial presentation of isolated congenital partial absence of the pericardium with similar physical examination and radiological findings. Copyright © 2016 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.

  15. Identification of immune genes and proteins involved in the response of bovine mammary tissue to Staphylococcus aureus infection

    PubMed Central

    Lutzow, Ylva C Strandberg; Donaldson, Laurelea; Gray, Christian P; Vuocolo, Tony; Pearson, Roger D; Reverter, Antonio; Byrne, Keren A; Sheehy, Paul A; Windon, Ross; Tellam, Ross L

    2008-01-01

    Background Mastitis in dairy cattle results from infection of mammary tissue by a range of micro-organisms but principally coliform bacteria and Gram positive bacteria such as Staphylococcus aureus. The former species are often acquired by environmental contamination while S. aureus is particularly problematic due to its resistance to antibiotic treatments and ability to reside within mammary tissue in a chronic, subclinical state. The transcriptional responses within bovine mammary epithelial tissue subjected to intramammary challenge with S. aureus are poorly characterised, particularly at the earliest stages of infection. Moreover, the effect of infection on the presence of bioactive innate immune proteins in milk is also unclear. The nature of these responses may determine the susceptibility of the tissue and its ability to resolve the infection. Results Transcriptional profiling was employed to measure changes in gene expression occurring in bovine mammary tissues sampled from three dairy cows after brief and graded intramammary challenges with S. aureus. These limited challenges had no significant effect on the expression pattern of the gene encoding β-casein but caused coordinated up-regulation of a number of cytokines and chemokines involved in pro-inflammatory responses. In addition, the enhanced expression of two genes, S100 calcium-binding protein A12 (S100A12) and Pentraxin-3 (PTX3) corresponded with significantly increased levels of their proteins in milk from infected udders. Both genes were shown to be expressed by mammary epithelial cells grown in culture after stimulation with lipopolysaccharide. There was also a strong correlation between somatic cell count, a widely used measure of mastitis, and the level of S100A12 in milk from a herd of dairy cows. Recombinant S100A12 inhibited growth of Escherichia coli in vitro and recombinant PTX3 bound to E. coli as well as C1q, a subunit of the first component of the complement cascade. Conclusion The

  16. Fresh autologous pericardium for leaflet perforation repair in mitral valve infective endocarditis.

    PubMed

    Evans, Charles F; DeFilippi, Christopher R; Shang, Eric; Griffith, Bartley P; Gammie, James S

    2013-07-01

    There is clear evidence that mitral valve (MV) repair is superior to replacement for MV infective endocarditis (IE). Leaflet perforation is a common pathologic finding in MV IE, and leaflet patch repair with glutaraldehyde-treated autologous or bovine pericardium is the currently accepted method of MV repair. In the present study, fresh autologous pericardium (FAP) was used universally to treat leaflet perforation in MV IE, and the mid-term clinical and echocardiographic outcomes were determined. Between 2002 and 2009, a total of 20 patients with leaflet perforations from MV IE underwent patch repair with FAP. Follow up echocardiography was performed in a core laboratory. There was one operative death (5%) secondary to sepsis, and three late deaths (15%). Late echocardiograms were available for review from 16 of the 19 patients (84%) who survived surgery. The mean time to follow up echocardiography was 793 +/- 663 days. The mitral regurgitation (MR) grade was mild or less in 14/16 patients (88%), moderate in one patient (6%), and severe in one patient (6%). The mean gradient was 4.8 +/- 2.7 mmHg, and the ejection fraction was preserved in all patients (63 +/- 4%). No expansion, retraction or calcification of the patches was observed. Freedom from reoperation, reinfection and thromboembolism was 100%. Fresh autologous pericardium for MV leaflet patch repair in IE is associated with good mid-term valve function. Given the association between late calcification and the glutaraldehyde treatment of bioprosthetic valves and this favorable experience, it is believed that FAP is an acceptable alternative for leaflet repair in MV IE.

  17. Determining the modulus of intact bovine vertebral cancellous bone tissue: Development and validation of a protocol

    NASA Astrophysics Data System (ADS)

    Engbretson, Andrew Craig

    Cancellous, or spongy, bone accounts for nearly 80% of the human skeleton's internal surface area, despite comprising only 20% of its mass. It is made up of a network of struts and plates that provide lightweight internal support to mammalian bones. In addition, it often serves as the main interface between the skeletal system and implanted devices such as artificial hips, knees, and fracture fixation devices. However, hip arthroplasties can succumb to loosening of the implant due to bone resorption, which is thought to be caused by a mismatch in both apparent and real stiffness between the device and the surrounding bone. Many studies have attempted to determine the Young's modulus of cancellous bone tissue, but the results are far from being in agreement. Reported values range from less than 1 to nearly 20 GPa. In addition, the small size of trabeculae has made dissection and testing a challenge. In this thesis, whole individual trabeculae from a bovine lumbar spine were tested in three-point bending to determine their Young's modulus using custom-made equipment to fit a miniature single-axis testing device. The device itself was validated by testing materials with moduli ranging from 1 to 200 GPa. The structure of the cancellous bone and the morphology of the individual struts were determined using micro x-ray computed tomography (muXCT). Individual struts were manually isolated from slices made using a low-speed saw under constant lubrication and measured under a stereomicroscope. Samples exhibiting no machined surfaces (and thus deemed to be whole, or "uncut" were compared to struts that had been cut by the saw during sectioning. Validation showed that the system was capable of determining the modulus of materials that were approximately five times stiffer than the expected cancellous modulus (copper, at 115 GPa) to within 10% of published values. This gave confidence in the results for bone. The modulus of the "uncut" specimens was found to be 15.28 2.26 GPa

  18. Pharmacokinetics of tildipirosin in bovine plasma, lung tissue, and bronchial fluid (from live, nonanesthetized cattle).

    PubMed

    Menge, M; Rose, M; Bohland, C; Zschiesche, E; Kilp, S; Metz, W; Allan, M; Röpke, R; Nürnberger, M

    2012-12-01

    The pharmacokinetics of tildipirosin (Zuprevo(®) 180 mg/mL solution for injection for cattle), a novel 16-membered macrolide for treatment, control, and prevention of bovine respiratory disease, were investigated in studies collecting blood plasma, lung tissue, and in vivo samples of bronchial fluid (BF) from cattle. After single subcutaneous (s.c.) injection at 4 mg/kg body weight, maximum plasma concentration (C(max)) was 0.7 μg/mL. T(max) was 23 min. Mean residence time from the time of dosing to the time of last measurable concentration (MRT(last)) and terminal half-life (T(1/2) ) was 6 and 9 days, respectively. A strong dose-response relationship with no significant sex effect was shown for both C(max) and area under the plasma concentration-time curve from time 0 to the last sampling time with a quantifiable drug concentration (AUC(last) ) over the range of doses up to 6 mg/kg. Absolute bioavailability was 78.9%. The volume of distribution based on the terminal phase (V(z)) was 49.4 L/kg, and the plasma clearance was 144 mL/h/kg. The time-concentration profile of tildipirosin in BF and lung far exceeded those in blood plasma. In lung, tildipirosin concentrations reached 9.2 μg/g at 4 h, peaked at 14.8 μg/g at day 1, and slowly declined to 2.0 μg/g at day 28. In BF, the concentration of tildipirosin reached 1.5 and 3.0 μg/g at 4 and 10 h, maintained a plateau of about 3.5 μg/g between day 1 and 3, and slowly declined to 1.0 at day 21. T(1/2) in lung and BF was approximately 10 and 11 days. Tildipirosin is rapidly and extensively distributed to the respiratory tract followed by slow elimination.

  19. Design and Development of Potential Tissue Engineering Scaffolds from Structurally Different Longitudinal Parts of a Bovine-Femur

    PubMed Central

    Pramanik, Sumit; Pingguan-Murphy, Belinda; Cho, Jongman; Osman, Noor Azuan Abu

    2014-01-01

    The complex architecture of the cortical part of the bovine-femur was examined to develop potential tissue engineering (TE) scaffolds. Weight-change and X-ray diffraction (XRD) results show that significant phase transformation and morphology conversion of the bone occur at 500–750°C and 750–900°C, respectively. Another breakthrough finding was achieved by determining a sintering condition for the nucleation of hydroxyapatite crystal from bovine bone via XRD technique. Scanning electron microscopy results of morphological growth suggests that the concentration of polymer fibrils increases (or decreases, in case of apatite crystals) from the distal to proximal end of the femur. Energy-dispersive analysis of X-ray, Fourier transform infrared, micro-computer tomography, and mechanical studies of the actual composition also strongly support our microscopic results and firmly indicate the functionally graded material properties of bovine-femur. Bones sintered at 900 and 1000°C show potential properties for soft and hard TE applications, respectively. PMID:25068570

  20. Direct polymerase chain reaction from blood and tissue samples for rapid diagnosis of bovine leukemia virus infection.

    PubMed

    Nishimori, Asami; Konnai, Satoru; Ikebuchi, Ryoyo; Okagawa, Tomohiro; Nakahara, Ayako; Murata, Shiro; Ohashi, Kazuhiko

    2016-06-01

    Bovine leukemia virus (BLV) infection induces bovine leukemia in cattle and causes significant financial harm to farmers and farm management. There is no effective therapy or vaccine; thus, the diagnosis and elimination of BLV-infected cattle are the most effective method to eradicate the infection. Clinical veterinarians need a simpler and more rapid method of diagnosing infection, because both nested polymerase chain reaction (PCR) and real-time PCR are labor intensive, time-consuming, and require specialized molecular biology techniques and expensive equipment. In this study, we describe a novel PCR method for amplifying the BLV provirus from whole blood, thus eliminating the need for DNA extraction. Although the sensitivity of PCR directly from whole blood (PCR-DB) samples as measured in bovine blood containing BLV-infected cell lines was lower than that of nested PCR, the PCR-DB technique showed high specificity and reproducibility. Among 225 clinical samples, 49 samples were positive by nested PCR, and 37 samples were positive by PCR-DB. There were no false positive samples; thus, PCR-DB sensitivity and specificity were 75.51% and 100%, respectively. However, the provirus loads of the samples detected by nested PCR and not PCR-DB were quite low. Moreover, PCR-DB also stably amplified the BLV provirus from tumor tissue samples. PCR-DB method exhibited good reproducibility and excellent specificity and is suitable for screening of thousands of cattle, thus serving as a viable alternative to nested PCR and real-time PCR.

  1. Altered microRNA expression in bovine subcutaneous and visceral adipose tissues from cattle under different diet.

    PubMed

    Romao, Josue Moura; Jin, Weiwu; He, Maolong; McAllister, Tim; Guan, Le Luo

    2012-01-01

    MicroRNAs (miRNAs) are a class of molecular regulators found to participate in numerous biological processes, including adipogenesis in mammals. This study aimed to evaluate the differences of miRNA expression between bovine subcutaneous (backfat) and visceral fat depots (perirenal fat) and the dietary effect on miRNA expression in these fat tissues. Fat tissues were collected from 16 Hereford×Aberdeen Angus cross bred steers (15.5 month old) fed a high-fat diet (5.85% fat, n = 8) or control diet (1.95% fat, n = 8). Total RNA from each animal was subjected to miRNA microarray analysis using a customized Agilent miRNA microarray containing 672 bovine miRNA probes. Expression of miRNAs was not equal between fat depots as well as diets: 207 miRNAs were detected in both fat depots, while 37 of these were found to be tissue specific; and 169 miRNAs were commonly expressed under two diets while 75 were diet specific. The number of miRNAs detected per animal fed the high fat diet was higher than those fed control diet (p = 0.037 in subcutaneous fat and p = 0.002 visceral fat). Further qRT-PCR analysis confirmed that the expression of some miRNAs was highly influenced by diet (miR-19a, -92a, -92b, -101, -103, -106, -142-5p, and 296) or fat depot (miR-196a and -2454). Our results revealed that the miRNA may differ among adipose depots and level of fat in the diet, suggesting that miRNAs may play a role in the regulation of bovine adipogenesis.

  2. Detection of PrP(Sc) in peripheral tissues of clinically affected cattle after oral challenge with bovine spongiform encephalopathy.

    PubMed

    Franz, Martin; Eiden, Martin; Balkema-Buschmann, Anne; Greenlee, Justin; Schatzl, Hermann; Fast, Christine; Richt, Jürgen; Hildebrandt, Jan-Peter; Groschup, Martin H

    2012-12-01

    Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative prion disease that mainly affects cattle. Transmission of BSE to humans caused a variant form of Creutzfeldt-Jakob disease. Following infection, the protease-resistant, disease-associated isoform of prion protein (PrP(Sc)) accumulates in the central nervous system and in other tissues. Many countries have defined bovine tissues that may contain prions as specified risk materials, which must not enter the human or animal food chains and therefore must be discarded. Ultrasensitive techniques such as protein misfolding cyclic amplification (PMCA) have been developed to detect PrP(Sc) when present in minuscule amounts that are not readily detected by other diagnostic methods such as immunohistochemistry or Western blotting. This study was conducted to determine when and where PrP(Sc) can be found by PMCA in cattle orally challenged with BSE. A total of 48 different tissue samples from four cattle infected orally with BSE at various clinical stages of disease were examined using a standardized PMCA protocol. The protocol used brain homogenate from bovine PrP transgenic mice (Tgbov XV) as substrate and three consecutive rounds of PMCA. Using this protocol, PrP(Sc) was found in the brain, spinal cord, nerve ganglia, optic nerve and Peyer's patches. The presence of PrP(Sc) was confirmed in adrenal glands, as well as in mesenteric lymph nodes - a finding that was reported recently by another group. Interestingly, additional positive results were obtained for the first time in the oesophagus, abomasum, rumen and rectum of clinically affected cattle.

  3. Characterization of an apically derived epithelial membrane glycoprotein from bovine milk, which is expressed in capillary endothelia in diverse tissues

    PubMed Central

    1985-01-01

    A glycoprotein (PAS IV) of apparent Mr 76,000 was purified from bovine milk-fat-globule membrane and partially characterized. PAS IV contained mannose, galactose, and sialic acid as principal sugars (approximately 5.3% total carbohydrate [wt/wt]) and existed in milk in at least four isoelectric variants. The glycoprotein appeared to be an integral membrane protein by several criteria. PAS IV was recovered in the detergent phase of Triton X-114 extracts of milk-fat-globule membrane at room temperature. When bound to membrane, PAS IV was resistant to digestion by a number of proteinases, although after solubilization with non-ionic detergents, the protein was readily degraded. Amino acid analysis of the purified protein revealed a high percentage of amino acids with nonpolar residues. The location of PAS IV was determined in bovine tissues by using immunofluorescence techniques. In mammary tissue, PAS IV was located on both the apical surfaces of secretory epithelial cells and endothelial cells of capillaries. This glycoprotein was also detected in endothelial cells of heart, liver, spleen, pancreas, salivary gland, and small intestine. In addition to mammary epithelial cells, PAS IV was also located in certain other epithelial cells, most notably the bronchiolar epithelial cells of lung. The potential usefulness of this protein as a specific marker of capillary endothelial cells in certain tissues is discussed. PMID:3881456

  4. Neither bovine somatotropin nor growth hormone-releasing factor alters expression of thyroid hormone receptors in liver and mammary tissues.

    PubMed

    Capuco, A V; Binelli, M; Tucker, H A

    2011-10-01

    Physiological effects of thyroid hormones are mediated primarily by binding of triiodothyronine to specific nuclear receptors. Organ-specific changes in production of triiodothyronine from its prohormone, thyroxine, have been hypothesized to target the action of thyroid hormones on the mammary gland and play a role in mediating or augmenting a galactopoietic response to bovine somatotropin (bST). Additionally, tissue responsiveness to thyroid hormones may be altered by changes in the number or affinity of nuclear receptors for thyroid hormones. In the present study, effects of bST and bovine growth hormone-releasing factor (bGRF) on thyroid hormone receptors in liver and mammary gland were studied. Lactating Holstein cows received continuous infusions of bST or bGRF for 63 d or served as uninfused controls. Nuclei were isolated from harvested mammary and liver tissues and incubated with [(125)I]-triiodothyronine. Treatments did not alter the capacity or affinity of specific binding sites for triiodothyronine in liver or mammary nuclei. Evaluation of transcript abundance for thyroid hormone receptors showed that isoforms of thyroid hormone receptor or retinoid receptor (which may influence thyroid receptor action) expressed in the mammary gland were not altered by bST or bGRF treatment. Data do not support the hypothesis that administration of bST or bGRF alters sensitivity of mammary tissue by changing expression of thyroid hormone receptors.

  5. Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization

    PubMed Central

    Albrecht, Elke; Komolka, Katrin; Kuzinski, Judith; Maak, Steffen

    2012-01-01

    Beside its role in melanogenesis, the agouti signaling protein (ASIP) has been related to obesity. The potentially crucial role in adipocyte development makes it a tempting candidate for economic relevant, fat related traits in farm animals. The objective of our study was to characterize the mRNA expression of different ASIP transcripts and of putative targets in different bovine tissues, as well as to study consequences on protein abundance and localization. ASIP mRNA abundance was determined by RT-qPCR in adipose and further tissues of cattle representing different breeds and crosses. ASIP mRNA was up-regulated more than 9-fold in intramuscular fat of Japanese Black cattle compared to Holstein (p<0.001). Further analyses revealed that a transposon-derived transcript was solely responsible for the increased ASIP mRNA abundance. This transcript was observed in single individuals of different breeds indicating a wide spread occurrence of this insertion at the ASIP locus in cattle. The protein was detected in different adipose tissues, skin, lung and liver, but not in skeletal muscle by Western blot with a bovine-specific ASIP antibody. However, the protein abundance was not related to the observed ASIP mRNA over-expression. Immuno-histochemical analyses revealed a putative nuclear localization of ASIP additionally to the expected cytosolic signal in different cell types. The expression of melanocortin receptors (MCR) 1 to 5 as potential targets for ASIP was analyzed by RT-PCR in subcutaneous fat. Only MC1R and MC4R were detected indicating a similar receptor expression like in human adipose tissue. Our results provide evidence for a widespread expression of ASIP in bovine tissues at mRNA and, for the first time, at protein level. ASIP protein is detectable in adipocytes as well as in further cells of adipose tissue. We generated a basis for a more detailed investigation of ASIP function in peripheral tissues of various mammalian species. PMID:22530003

  6. Complete genome sequences of two bovine viral diarrhea viruses isolated from brain tissues of nonambulatory (downer) cattle.

    PubMed

    Oem, Jae-Ku; Joo, Soo-Kyung; An, Dong-Jun

    2013-09-12

    Here, we report the complete genome sequences of two bovine viral diarrhea viruses (BVDVs) (strains 11F011 and 12F004) isolated from brain tissues from nonambulatory (downer) cattle. The complete genomes of strains 11F011 and 12F004 contain 12,287 nucleotides (nt) with a single large open reading frame and 12,301 nt with a single large open reading frame, respectively. Phylogenetic analysis indicated that these strains belong to the BVDV-2a and -1b genotypes, respectively.

  7. Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold

    PubMed Central

    Feng, Yuping; Wang, Jiao; Ling, Shixin; Li, Zhuo; Li, Mingsheng; Li, Qiongyi; Ma, Zongren; Yu, Sijiu

    2014-01-01

    The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells following induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined specific neuronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuronal-specific proteins, including βIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differentiation medium differentiated into a multilayered neural network-like structure with long nerve fibers that was composed of several parallel microfibers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sectioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve. PMID:25598779

  8. X-ray fluorescence-based differentiation of neck tissues in a bovine model: implications for potential intraoperative use.

    PubMed

    Lahav, G; Shilstein, S; Shchemelinin, S; Ikher, S; Halperin, D; Chechik, R; Breskin, A

    2015-05-01

    This study explores the possibility of using X-ray fluorescence (XRF)-based trace-element analysis for differentiation of various bovine neck tissues. It is motivated by the requirement for an intra-operative in-vivo method for identifying parathyroid glands, particularly beneficial in surgery in the central neck-compartment. Using a dedicated X-ray spectral analysis, we examined ex-vivo XRF spectra from various histologically verified fresh neck tissues from cow, which was chosen as the animal model; these tissues included fat, muscle, thyroid, parathyroid, lymph nodes, thymus and salivary gland. The data for six trace elements K, Fe, Zn, Br, Rb and I, provided the basis for tissue identification by using multi-parameter analysis of the recorded XRF spectra. It is shown that the combination of XRF signals from these elements is sufficient for a reliable tissue differentiation. The average total abundance of these trace elements was evaluated in each tissue type, including parathyroid and salivary gland for the first time. It is shown that some tissues can unequivocally be identified on the basis of the abundance of a single element, for example, iodine and zinc for the identification of thyroid gland and muscle, respectively.

  9. Congenital defects of pericardium: case reports and review of literature.

    PubMed

    Cuccuini, Matteo; Lisi, Francesca; Consoli, Arturo; Mancini, Sara; Bellino, Valentina; Galanti, Giorgio; Capaccioli, Leonardo

    2013-01-01

    Pericardial defects are a rare condition, generally asymptomatic, due to failure in development of pericardial sac. They are difficult to detect, particularly the complete absence of the pericardium. At present magnetic resonance imaging (MRI) is considered the best performing exam and it could be the first choice in the diagnosis of agenesia of the pericardium. A comprehensive review of dedicated books and PubMed literature was performed and three clinical cases have been analyzed. We report three cases, one with partial and two with total agenesia of the left pericardium, seen at our Institute and diagnosed at MRI as unexpected findings. The diagnosis of a congenital defect of the pericardium is challenging, even for expert radiologists, because this kind of deformity does not provide specific clinical nor conventional radiology findings. Nowadays, the most effective instrument for a certain diagnosis is MRI, interlocked with electrocardiography.

  10. Effects of N-acetylimidazole on oxytocin binding in bovine mammary tissue

    SciTech Connect

    Zhao, X.; Gorewit, R.C.; Currie, W.B. )

    1990-01-01

    The effects of N-acetylimidazole on specific binding of oxytocin to microsomal fractions of bovine mammary gland were studied. N-acetylimidazole suppressed oxytocin binding, with time and concentration dependence. Decreased oxytocin binding activity appeared to be due to decreased affinity of the hormone for its receptor. Acetylation of oxytocin, rather than of oxytocin receptors, seemed to be responsible for the decreased binding.

  11. A fluorescence based method, exploiting lipofuscin, for the real-time detection of central nervous system (CNS) tissues on bovine carcasses

    USDA-ARS?s Scientific Manuscript database

    The removal of Central Nervous System (CNS) tissues as part of Bovine Spongiform Encephalopathy (BSE) risk material is one of the highest priority tasks to avoid contamination of the human food chain with BSE. No currently available method enables the real-time detection of possible CNS tissue conta...

  12. Closure of pericardium after open heart surgery. A way to prevent postoperative cardiac tamponade.

    PubMed Central

    Nandi, P; Leung, J S; Cheung, K L

    1976-01-01

    Between July 1968 and December 1975, 821 patients underwent open heart operations. In 596 cases the pericardium was left open and in 225 the pericardium was closed. Forty-one patients in the open pericardium group required reoperation and 23 of these had tamponade. Four patients in the closed pericardium group had reoperation but there was not a single case of tamponade. In most cases that required reoperation the bleeding was from extrapericardial sources. Absence of tamponade in the closed pericardium group can be explained by the fact that blood from extrapericardial sources of bleeding cannot collect round the heart because the pericardium is closed. Thus closure of pericardium helps to prevent tamponade. Reoperations some months or years after the original operation are technically easier and less hazardous if the pericardium has been closed because the closed pericardium prevents the heart from becoming adherent to the back of sternum and also because there are fewer adhesions in the pericardial cavity. PMID:795444

  13. Flavin reductase: sequence of cDNA from bovine liver and tissue distribution.

    PubMed Central

    Quandt, K S; Hultquist, D E

    1994-01-01

    Flavin reductase catalyzes electron transfer from reduced pyridine nucleotides to methylene blue or riboflavin, and this catalysis is the basis of the therapeutic use of methylene blue or riboflavin in the treatment of methemoglobinemia. A cDNA for a mammalian flavin reductase has been isolated and sequenced. Degenerate oligonucleotides, with sequences based on amino acid sequences of peptides derived from bovine erythrocyte flavin reductase, were used as primers in PCR to selectively amplify a partial cDNA that encodes the bovine reductase. The template used in the PCR was first strand cDNA synthesized from bovine liver total RNA using oligo(dT) primers. A PCR product was used as a specific probe to screen a bovine liver cDNA library. The sequence determined from two overlapping clones contains an open reading frame of 621 nucleotides and encodes 206 amino acids. The amino acid sequence deduced from the bovine liver flavin reductase cDNA matches the amino acid sequences determined for erythrocyte reductase-derived peptides, and the predicted molecular mass of 22,001 Da for the liver reductase agrees well with the molecular mass of 21,994 Da determined for the erythrocyte reductase by electrospray mass spectrometry. The amino acid sequence at the N terminus of the reductase has homology to sequences of pyridine nucleotide-dependent enzymes, and the predicted secondary structure, beta alpha beta, resembles the common nucleotide-binding structural motif. RNA blot analysis indicates a single 1-kilobase reductase transcript in human heart, kidney, liver, lung, pancreas, placenta, and skeletal muscle. Images PMID:7937764

  14. The telescoping suture--Part II: A novel method to improve the mechanical behavior of a new biomaterial: ostrich pericardium.

    PubMed

    García Páez, J M; Jorge Herrero, E; Rocha, A; Martín-Maestro, M; Castillo-Olivares, J L; Millán, I; Carrera Sanmartín, A; Cordón, A

    2002-10-01

    Ostrich pericardium, sutured using a telescoping or overlapping technique, was studied to determine its mechanical behavior. From each of 12 pericardial sacs, four contiguous strips were cut longitudinally, from root to apex, and another four contiguous strips were cut in transverse direction. One of the strips in each set of four was used as an unsutured control and the remaining three were sutured by overlapping 0.5 cm of the tissue and sewing with Gore-tex, Prolene or Pronova. These 96 samples were then subjected to tensile testing along their major axes until rupture. The tensile stresses recorded in the suture materials at the moment tears appeared in the pericardium ranged between 55.99 MPa and 70.23 MPa for Gore-tex in samples cut in the two directions. Shear stress became ostensible at 56 MPa, with clearly evident tears. However, microfracture of the collagen fibers must be produced at much lower stress levels. The comparison of the resistance in kilograms (machine-imposed), without taking into account the sections in which the load was applied, demonstrated only a slight loss of load when the telescoping suture was employed in ostrich pericardium samples. Ostrich pericardium may continue to be an alternative biological material for the construction of heart valve leaflets.

  15. Feasibility of pig and human-derived aortic valve interstitial cells seeding on fixative-free decellularized animal pericardium.

    PubMed

    Santoro, Rosaria; Consolo, Filippo; Spiccia, Marco; Piola, Marco; Kassem, Samer; Prandi, Francesca; Vinci, Maria Cristina; Forti, Elisa; Polvani, Gianluca; Fiore, Gianfranco Beniamino; Soncini, Monica; Pesce, Maurizio

    2016-02-01

    Glutaraldehyde-fixed pericardium of animal origin is the elective material for the fabrication of bio-prosthetic valves for surgical replacement of insufficient/stenotic cardiac valves. However, the pericardial tissue employed to this aim undergoes severe calcification due to chronic inflammation resulting from a non-complete immunological compatibility of the animal-derived pericardial tissue resulting from failure to remove animal-derived xeno-antigens. In the mid/long-term, this leads to structural deterioration, mechanical failure, and prosthesis leaflets rupture, with consequent need for re-intervention. In the search for novel procedures to maximize biological compatibility of the pericardial tissue into immunocompetent background, we have recently devised a procedure to decellularize the human pericardium as an alternative to fixation with aldehydes. In the present contribution, we used this procedure to derive sheets of decellularized pig pericardium. The decellularized tissue was first tested for the presence of 1,3 α-galactose (αGal), one of the main xenoantigens involved in prosthetic valve rejection, as well as for mechanical tensile behavior and distensibility, and finally seeded with pig- and human-derived aortic valve interstitial cells. We demonstrate that the decellularization procedure removed the αGAL antigen, maintained the mechanical characteristics of the native pig pericardium, and ensured an efficient surface colonization of the tissue by animal- and human-derived aortic valve interstitial cells. This establishes, for the first time, the feasibility of fixative-free pericardial tissue seeding with valve competent cells for derivation of tissue engineered heart valve leaflets. © 2015 Wiley Periodicals, Inc.

  16. Tensile strength of human pericardium treated with glutaraldehyde.

    PubMed

    Yamashita, Hiromasa; Ozaki, Shigeyuki; Iwasaki, Kiyotaka; Kawase, Isamu; Nozawa, Yukinari; Umezu, Mitsuo

    2012-01-01

    We have reconstructed aortic valves using autologous pericardium treated with glutaraldehyde since April 2007. However, the strength of the human pericardium has not been confirmed. We compared tensile strength between glutaraldehyde-treated human pericardium and aortic valve leaflets with various degrees of calcification to determine their suitability for use in aortic valve reconstruction. We measured the ultimate tensile strength and elasticity of samples of glutaraldehyde-treated pericardia (n = 8), non-calcified (n = 12), calcified (n = 9) and decalcified (n = 21) aortic leaflets collected from 23 patients who underwent aortic valve surgery. Aortic valves were decalcified using a cavitational ultrasonic surgical aspirator. The pericardium was immersed in 0.6% buffered glutaraldehyde for 10 minutes and then rinsed three times for 6 minutes each in normal saline. The ultimate tensile strength of the glutaraldehyde-treated human pericardium, non-calcified, calcified and decalcified leaflets was 10, 2.8, 1.0 and 0.8 MPa, respectively. The ultimate tensile strength of glutaraldehyde-treated human pericardium was 4 times higher than non-calcified leaflets, indicating its suitability for application to aortic valve reconstruction. Calcified leaflets were slightly stronger than decalcified leaflets. Thus, calcification can be removed without altering the tensile strength of valve materials.

  17. Expression, localization, and functional model of cholesterol transporters in lactating and nonlactating mammary tissues of murine, bovine, and human origin.

    PubMed

    Mani, Orlando; Körner, Meike; Sorensen, Martin T; Sejrsen, Kristen; Wotzkow, Carlos; Ontsouka, Corneille E; Friis, Robert R; Bruckmaier, Rupert M; Albrecht, Christiane

    2010-08-01

    Members of the ATP-binding cassette (ABC) transporters play a pivotal role in cellular lipid efflux. To identify candidate cholesterol transporters implicated in lipid homeostasis and mammary gland (MG) physiology, we compared expression and localization of ABCA1, ABCG1, and ABCA7 and their regulatory genes in mammary tissues of different species during the pregnancy-lactation cycle. Murine and bovine mammary glands (MGs) were investigated during different functional stages. The abundance of mRNAs was determined by quantitative RT-PCR. Furthermore, transporter proteins were localized in murine, bovine, and human MGs by immunohistochemistry. In the murine MG, ABCA1 mRNA abundance was elevated during nonlactating compared with lactating stages, whereas ABCA7 and ABCA1 mRNA profiles were not altered. In the bovine MG, ABCA1, ABCG1, and ABCA7 mRNAs abundances were increased during nonlactating stages compared with lactation. Furthermore, associations between mRNA levels of transporters and their regulatory genes LXRalpha, PPARgamma, and SREBPs were found. ABCA1, ABCG1, and ABCA7 proteins were localized in glandular MG epithelial cells (MEC) during lactation, whereas during nonlactating stages, depending on species, the proteins showed distinct localization patterns in MEC and adipocytes. Our results demonstrate that ABCA1, ABCG1, and ABCA7 are differentially expressed between lactation and nonlactating stages and in association with regulatory genes. Combined expression and localization data suggest that the selected cholesterol transporters are universal MG transporters involved in transport and storage of cholesterol and in lipid homeostasis of MEC. Because of the species-specific expression patterns of transporters in mammary tissue, mechanisms of cholesterol homeostasis seem to be differentially regulated between species.

  18. Survival and Diversity of Human Homologous Dietary MicroRNAs in Conventionally Cooked Top Sirloin and Dried Bovine Tissue Extracts.

    PubMed

    Dever, Joseph T; Kemp, Michael Q; Thompson, Amber L; Keller, Hana G K; Waksmonski, James C; Scholl, Chris D; Barnes, David M

    2015-01-01

    Dietary microRNAs (miRNAs), notably those found in milk, are currently being investigated for their potential to elicit biological effects via canonical binding to human messenger RNA targets once ingested. Besides milk, beef and other bovine tissue-derived ingredients could also be a relevant source of potentially bioactive dietary miRNAs. In this study, we characterized the human homologous miRNA profiles in food-grade, bovine-sourced sirloin, heart and adrenal tissue (raw, cooked, and pasteurized, freeze-dried extracts) via deep-sequencing and quantitative reverse transcription PCR (RT-qPCR). A total of 198 human homologous miRNAs were detected at 10 or more normalized reads in all replicates (n = 3) of at least one preparation method. Tissue origin rather than preparation method was the major differentiating factor of miRNA profiles, and adrenal-based miRNA profiles were the most distinct. The ten most prevalent miRNAs in each tissue represented 71-93% of the total normalized counts for all annotated miRNAs. In cooked sirloin, the most abundant miRNAs were miR-10b-5p, (48.8% of total annotated miRNA reads) along with the muscle-specific miR-1 (24.1%) and miR-206 (4.8%). In dried heart extracts, miR-1 (17.0%), miR-100-5p (16.1%) and miR-99a-5p (11.0%) gave the highest normalized read counts. In dried adrenal extracts, miR-10b-5p (71.2%) was the most prominent followed by miR-143-3p (7.1%) and 146b-5p (3.7%). Sequencing results for five detected and two undetected miRNAs were successfully validated by RT-qPCR. We conclude that edible, bovine tissues contain unique profiles of human homologous dietary miRNAs that survive heat-based preparation methods.

  19. Survival and Diversity of Human Homologous Dietary MicroRNAs in Conventionally Cooked Top Sirloin and Dried Bovine Tissue Extracts

    PubMed Central

    Thompson, Amber L.; Keller, Hana G. K.; Waksmonski, James C.; Scholl, Chris D.

    2015-01-01

    Dietary microRNAs (miRNAs), notably those found in milk, are currently being investigated for their potential to elicit biological effects via canonical binding to human messenger RNA targets once ingested. Besides milk, beef and other bovine tissue-derived ingredients could also be a relevant source of potentially bioactive dietary miRNAs. In this study, we characterized the human homologous miRNA profiles in food-grade, bovine-sourced sirloin, heart and adrenal tissue (raw, cooked, and pasteurized, freeze-dried extracts) via deep-sequencing and quantitative reverse transcription PCR (RT-qPCR). A total of 198 human homologous miRNAs were detected at 10 or more normalized reads in all replicates (n = 3) of at least one preparation method. Tissue origin rather than preparation method was the major differentiating factor of miRNA profiles, and adrenal-based miRNA profiles were the most distinct. The ten most prevalent miRNAs in each tissue represented 71–93% of the total normalized counts for all annotated miRNAs. In cooked sirloin, the most abundant miRNAs were miR-10b-5p, (48.8% of total annotated miRNA reads) along with the muscle-specific miR-1 (24.1%) and miR-206 (4.8%). In dried heart extracts, miR-1 (17.0%), miR-100-5p (16.1%) and miR-99a-5p (11.0%) gave the highest normalized read counts. In dried adrenal extracts, miR-10b-5p (71.2%) was the most prominent followed by miR-143-3p (7.1%) and 146b-5p (3.7%). Sequencing results for five detected and two undetected miRNAs were successfully validated by RT-qPCR. We conclude that edible, bovine tissues contain unique profiles of human homologous dietary miRNAs that survive heat-based preparation methods. PMID:26394052

  20. Large-scale chromatofocusing-based method for isolating thymosin beta 4 and thymosin beta 9 from bovine tissues.

    PubMed

    Roboti, A; Livaniou, E; Evangelatos, G P; Tsoupras, G; Tsolas, O; Ithakissios, D S

    1994-12-02

    A large-scale method for the isolation of thymosin beta 4 (up to 120 mg) and thymosin beta 9 (up to 40 mg) from bovine lung (up to 2 kg) was developed. The isolation protocol included tissue homogenization in 0.4 M HClO4, centrifugation, solid-phase extraction through LiChroprep RP-18 material, chromatofocusing on polybuffer exchanger PBE 94-modified Sepharose and dialysis against water. The isolated products were characterized by analytical isoelectric focusing, reversed-phase HPLC, electrospray ionization mass spectrometry and amino acid analysis. The method developed is rapid and convenient, requires no expensive equipment and can be used for the isolation of thymosin beta 4 and homologous peptides from various animal tissues.

  1. Beta-carotene is incorporated or mobilized along with triglycerides in bovine adipose tissue in response to insulin or epinephrine.

    PubMed

    Arias, E; González, A; Shimada, A; Varela-Echavarria, A; Ruiz-López, F; During, A; Mora, O

    2009-02-01

    Pasture fed cattle ingest substantial amounts of beta-carotene (beta-C). Not all of the carotenoid compound is transformed into vitamin A, but the surplus is deposited in adipose tissue (AT). The mechanisms of beta-C incorporation and mobilization are unknown. Two experiments were conducted using explants from bovine AT cultured in vitro. First, beta-C incorporation by explants from three animals was examined with different beta-C concentrations (0, 1, 5 and 20 microm) and different times of incubation (every 5 h up to 25 h). The data showed a significant increase of beta-C concentration in explants only for 20 microm beta-C. Secondly, effects of insulin and epinephrine on beta-C and triglyceride (TG) contents of explants were studied. Explants from six animals were incubated with either hormone and 0 or 20 microm beta-C for 20 h. Both TG and beta-C contents were affected positively by insulin and negatively by epinephrine. Interestingly, changes in ratios of beta-C/TG (hormone vs. control) were similar (1.7 x 10(-3) and 1.8 x 10(-3)), respectively, for insulin and epinephrine, indicating that beta-C level is directly related to TG content. We also report the presence of mRNA for beta-C 15, 15' oxygenase in bovine AT. The in vitro culture system using explants from bovine AT is a promising model to investigate factors that might affect the accumulation and metabolism of beta-C.

  2. A highly conserved homologue of bovine neurocalcin in Drosophila melanogaster is a Ca(2+)-binding protein expressed in neuronal tissues.

    PubMed

    Teng, D H; Chen, C K; Hurley, J B

    1994-12-16

    Polymerase chain reaction was used to search for genes encoding recoverin-like proteins in Drosophila melanogaster. We identified a gene that codes for a cognate of bovine neurocalcin; hence, we have named it neurocalcin (nca). A cDNA of nca was isolated and sequenced. The deduced polypeptide product of the cDNA is 22 kDa in size, and its amino acid sequence is 88% identical to that of bovine neurocalcin. This deduced Drosophila neurocalcin (DrosNCa) protein has three putative EF-hands and has a sequence in its NH2 terminus required for fatty acylation. DrosNCa was expressed in Escherichia coli and subsequently purified by phenyl-Sepharose chromatography and Mono Q anion exchange fast protein liquid chromatography. This recombinant protein was capable of binding 45Ca2+ and exhibited Ca(2+)-dependent mobility shifts in both SDS-polyacrylamide gel electrophoresis and native gel electrophoresis. DrosNCa was tritiated when it was coexpressed in E. coli with N-myristoyl transferase in the presence of [3H]myristic acid. The nca transcript was approximately 1 kilobase long, and tissue in situ hybridization showed that this message was present in the brain of adult flies. Antibodies raised against recombinant DrosNCa cross-reacted with rat hippocalcin on an immunoblot but not with bovine recoverin. When immunohistochemical analysis was performed, staining was observed throughout the central nervous system of adult flies, particularly in the neuropil, where neurons synapse. The nca locus maps to or near 76F on the Drosophila third chromosome.

  3. Quasispecies of bovine enteric and respiratory coronaviruses based on complete genome sequences and genetic changes after tissue culture adaptation.

    PubMed

    Zhang, Xinsheng; Hasoksuz, Mustafa; Spiro, David; Halpin, Rebecca; Wang, Shiliang; Vlasova, Anastasia; Janies, Daniel; Jones, Leandro R; Ghedin, Elodie; Saif, Linda J

    2007-06-20

    The genetic diversity of 2 pairs (AH65 and AH187) of wild type bovine coronaviruses (BCoV) sequenced directly from nasal (respiratory) and rectal (enteric) swabs of two feedlot calves with respiratory and enteric symptoms [Hasoksuz, M., Sreevatsan, S., Cho, K.O., Hoet, A.E., Saif, L.J., 2002b. Molecular analysis of the S1 subunit of the spike glycoprotein of respiratory and enteric bovine coronavirus isolates. Virus Res. 84 (1-2), 101-109.]. was analyzed. Sequence analysis of the complete genomes revealed differences at 123 and 149 nucleotides (nt) throughout the entire genome between the respiratory and enteric strains for samples AH65 and AH187, respectively, indicating the presence of intra-host BCoV quasispecies. In addition, significant numbers of sequence ambiguities were found in the genomes of some BCoV-R and BCoV-E strains, suggesting intra-isolate quasispecies. The tissue culture (TC) passaged counterparts of AH65 respiratory BCoV (AH65-R-TC) and enteric BCoV (AH65-E-TC) were also sequenced after 14 and 15 passages and 1 plaque purification in human rectal tumor cells (HRT-18), respectively. Compared to the parental wild type strains, tissue culture passage generated 104 nt changes in the AH65-E-TC isolate but only 8 nt changes in the AH65-R-TC isolate. Particularly noteworthy, the majority of nucleotide changes in the AH65-E-TC isolate occurred at the identical positions as the mutations occurring in the AH65-R strain from the same animal. These data suggest that BCoV evolves through quasispecies development, and that enteric BCoV isolates are more prone to genetic changes and may mutate to resemble respiratory BCoV strains after tissue culture passage.

  4. Hypoelastic Soft Tissues

    PubMed Central

    Freed, Alan D.; Einstein, Daniel R.; Sacks, Michael S.

    2010-01-01

    In Part I, a novel hypoelastic framework for soft-tissues was presented. One of the hallmarks of this new theory is that the well-known exponential behavior of soft-tissues arises consistently and spontaneously from the integration of a rate based formulation. In Part II, we examine the application of this framework to the problem of biaxial kinematics, which are common in experimental soft-tissue characterization. We confine our attention to an isotropic formulation in order to highlight the distinction between non-linearity and anisotropy. In order to provide a sound foundation for the membrane extension of our earlier hypoelastic framework, the kinematics and kinetics of in-plane biaxial extension are revisited, and some enhancements are provided. Specifically, the conventional stress-to-traction mapping for this boundary value problem is shown to violate the conservation of angular momentum. In response, we provide a corrected mapping. In addition, a novel means for applying loads to in-plane biaxial experiments is proposed. An isotropic, isochoric, hypoelastic, constitutive model is applied to an in-plane biaxial experiment done on glutaraldehyde treated bovine pericardium. The experiment is comprised of eight protocols that radially probe the biaxial plane. Considering its simplicity (two adjustable parameters) the model does a reasonably good job of describing the non-linear normal responses observed in these experimental data, which are more prevalent than are the anisotropic responses exhibited by this tissue. PMID:21394222

  5. Decellularization of bovine anterior cruciate ligament tissues minimizes immunogenic reactions to alpha-gal epitopes by human peripheral blood mononuclear cells.

    PubMed

    Yoshida, Ryu; Vavken, Patrick; Murray, Martha M

    2012-10-01

    Rupture of ACL is a common injury. While the current surgical treatments are effective, many patients still suffer from precocious osteoarthritis, and there is an increasing interest in bioengineering approaches to improve ACL repair. Bovine collagen is a material currently in use for tissue engineering of ligaments. The alpha-gal epitopes found on bovine cells are a source of immunogenic stimulus for human cells. In this study, we wished to determine if those epitopes could be removed sufficiently to mitigate an immunogenic response using either a decellularization protocol or decellularization followed by alpha-galactosidase treatment. Bovine ACLs were treated with Triton-X, sodium deoxycholate, ribonuclease, and deoxyribonuclease to remove cells. A subset of the decellularized tissues was further treated with alpha-galactosidase. Human peripheral blood mononuclear cells (PBMCs) were exposed to untreated, decellularized, and alpha-galactosidase-treated tissues, and PBMC migration and IL-6 release were measured. PBMCs were significantly more attracted to untreated ACL compared to decellularized or alpha-galactosidase-treated tissue, but no difference was seen between the two treatment groups. PBMCs also released significantly more IL-6 when exposed to untreated tissue compared to decellularized ACL or alpha-galactosidase-treated ACL, but no difference was seen between the two treatment groups. Immunohistochemistry using anti-alpha-gal antibody detected the epitopes throughout the untreated ACL, but similar areas of reaction were not seen on decellularized or alpha-galactosidase-treated ACL. These results suggest that our decellularization protocol minimizes the immunogenic reactions of human PBMCs to bovine ACL tissue. Therefore, decellularized bovine ACL tissue may be a safe, effective biomaterial for ACL injury treatments.

  6. Healing of human intrabony defects following regenerative periodontal therapy with a bovine-derived xenograft and guided tissue regeneration.

    PubMed

    Sculean, A; Stavropoulos, A; Windisch, P; Keglevich, T; Karring, T; Gera, I

    2004-06-01

    The purpose of the present study was to histologically evaluate the healing of human intrabony defects following treatment with either a bovine-derived xenograft (BDX) and guided tissue regeneration (GTR) [BDX + GTR] or a bovine-derived xenograft mixed with collagen (BDX Coll) and GTR [BDX Coll + GTR]. Eight patients with chronic periodontitis and each with one very deep intrabony defect around a tooth scheduled for extraction were treated with either a combination of BDX + GTR (five patients) or with BDX Coll + GTR (three patients). The postoperative healing was uneventful in all eight cases. After a healing period of 6 months, the teeth or roots were extracted together with some of their surrounding soft and hard tissues and subsequently fixed in 10% buffered formalin. Following decalcification in EDTA, the specimens were embedded in paraffin and 8-microm histological sections were cut in the mesio-distal direction, parallel to the long axes of the teeth. The sections were alternatively stained with hematoxylin and eosin, van Giesson's connective tissue stain or with the Ladevig's connective tissue staining method and examined under the light microscope. Generally, formation of new cementum with inserting collagen fibers was found in seven out of the eight treated cases, whereas in the remaining case (treated with BDX + GTR) the healing was characterized by formation of a long junctional epithelium along the debrided root surface and no formation of cementum or bone. In the specimens demonstrating periodontal regeneration the new cementum was always of a cellular type. In most cases, the graft particles were surrounded by bone. In some areas, the bone tissue around the graft particles was connected by perpendicularly inserting collagen fibers to the newly formed cementum on the root surface. The epithelium downgrowth stopped always at the most coronal part of the newly formed cementum. No remnants of the membrane material were observed in any of the biopsies

  7. Micromechanical Modeling Study of Mechanical Inhibition of Enzymatic Degradation of Collagen Tissues.

    PubMed

    Tonge, Theresa K; Ruberti, Jeffrey W; Nguyen, Thao D

    2015-12-15

    This study investigates how the collagen fiber structure influences the enzymatic degradation of collagen tissues. We developed a micromechanical model of a fibrous collagen tissue undergoing enzymatic degradation based on two central hypotheses. The collagen fibers are crimped in the undeformed configuration. Enzymatic degradation is an energy activated process and the activation energy is increased by the axial strain energy density of the fiber. We determined the intrinsic degradation rate and characteristic energy for mechanical inhibition from fibril-level degradation experiments and applied the parameters to predict the effect of the crimped fiber structure and fiber properties on the degradation of bovine cornea and pericardium tissues under controlled tension. We then applied the model to examine the effect of the tissue stress state on the rate of tissue degradation and the anisotropic fiber structures that developed from enzymatic degradation. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  8. Micromechanical Modeling Study of Mechanical Inhibition of Enzymatic Degradation of Collagen Tissues

    PubMed Central

    Tonge, Theresa K.; Ruberti, Jeffrey W.; Nguyen, Thao D.

    2015-01-01

    This study investigates how the collagen fiber structure influences the enzymatic degradation of collagen tissues. We developed a micromechanical model of a fibrous collagen tissue undergoing enzymatic degradation based on two central hypotheses. The collagen fibers are crimped in the undeformed configuration. Enzymatic degradation is an energy activated process and the activation energy is increased by the axial strain energy density of the fiber. We determined the intrinsic degradation rate and characteristic energy for mechanical inhibition from fibril-level degradation experiments and applied the parameters to predict the effect of the crimped fiber structure and fiber properties on the degradation of bovine cornea and pericardium tissues under controlled tension. We then applied the model to examine the effect of the tissue stress state on the rate of tissue degradation and the anisotropic fiber structures that developed from enzymatic degradation. PMID:26682825

  9. Late results of mitral valve repair with glutaraldehyde-treated autologous pericardium.

    PubMed

    Shomura, Yu; Okada, Yukikatsu; Nasu, Michihiro; Koyama, Tadaaki; Yuzaki, Mitsuru; Murashita, Takashi; Fukunaga, Naoto; Konishi, Yasunobu

    2013-06-01

    Mitral valve repair is an established surgical procedure for treating severe organic mitral regurgitation. The mechanisms of mitral regurgitation due to infective endocarditis include rheumatic disease and congenital diseases such as a lack of leaflet tissue, and thus additional material is required to create a functional coaptation surface. We review our experience with 139 patients who underwent mitral valve repair with glutaraldehyde-treated autologous pericardium to treat organic mitral regurgitation between March 1992 and November 2011. Mitral valve disease mainly consisted of infective endocarditis in 51 patients (active, n = 32; healed, n = 19) and rheumatic disease in 47. This procedure was also applied to 12 patients who required reoperation after mitral valve repair for degenerative, congenital, or rheumatic mitral regurgitation. The mean follow-up was 4.5 ± 4.3 years (maximum 19.1). Actuarial survival at 10 years was 84% ± 5%. Eleven reoperations proceeded at a mean of 68 months after surgery. The causes of reoperation were rheumatic disease progression (n = 4), infection (n = 3), patch dehiscence (n = 2), progressive fibrosis of the remaining mitral valve tissue after infective endocarditis (n = 1), and patch tear (n = 1). Mitral valves were replaced in 8 patients and re-repaired in 3 patients. The autologous pericardium was not calcified at the time of reoperation. The rate of freedom from reoperation was 82% ± 7% at 10 years. Mitral valves that might otherwise require replacement can be durably and predictably repaired using glutaraldehyde-treated autologous pericardium. Copyright © 2013 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.

  10. Liquid chromatography/tandem mass spectrometry method to determine boldenone in bovine liver tissues.

    PubMed

    Granja, Rodrigo H M M; Salerno, Alessandro G; de Lima, Andreia C; Montalvo, Cynthia; Reche, Karine V G; Giannotti, Fabio M; Wanschel, Amarylis C B A

    2014-01-01

    Boldenone, an androgenic steroid, is forbidden for use in meat production in most countries worldwide. Residues of this drug in food present a potential risk to consumers. A sensitive LC/MS/MS method for analysis of 17β-boldenone using boldenone-d3 as an internal standard was developed. An enzymatic hydrolysis and extraction using ethyl acetate, methanol, and hexane were performed in the sample preparation. Parameters such as decision limit (CCα), detection capability (CCβ), precision, recovery, and ruggedness were evaluated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC) and International Organization for Standardization/International Electrotechnical Commission 17025:2005. CCα and CCβ were determined to be 0.17 and 0.29 μg/kg, respectively. Average recoveries from bovine liver samples fortified with 1, 1.5, and 2 μg/kg were around 100%. A complete statistical analysis was performed on the results obtained, including an estimation of the method uncertainty. The method is considered robust after being subjected to day-to-day analytical variations and has been used as a standard method in Brazil to report boldenone levels in bovine liver.

  11. Frozen and fresh ovarian tissue require different culture media to promote in vitro development of bovine preantral follicles.

    PubMed

    Castro, Simone Vieira; Carvalho, Adeline Andrade; Silva, Cleidson Manoel Gomes; Santos, Francielli Weber; Campello, Cláudio Cabral; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

    2014-10-01

    The aim of this study was to evaluate the efficiency of different media in the in vitro culture of bovine preantral follicles that were used either fresh or following slow freezing treatment. Frozen and fresh noncultured or cultured ovarian fragments were processed for histological, viability, and cell proliferation analyses. For cryopreservation, a solution containing 1.5 M ethylene glycol was frozen in a programmable biological freezer. After thawing, a portion of the samples was destined for frozen controls. The remainder were cultured in vitro for 5 days in three media: α-MEM, McCoy, or M199. Samples from these culture media were collected on days 1 and 5 for quantification of reactive oxygen species (ROS) and for hormonal assays. In fresh-cultured tissues, the percentage of morphologically normal follicles was significantly higher when cultured in M199 compared to that in the other media. In frozen-cultured tissues, McCoy medium was significantly superior to the other media, and was the only treatment that helped in maintaining the viability similar to fresh and frozen controls. Upon quantification of the nucleolus organizer region, we observed greater proliferation of granulosa cells in the frozen-cultured tissues with McCoy medium, and lesser proliferation in fresh-cultured tissues only with α-MEM. In frozen-cultured tissues, ROS levels were highest at day 1 and progressively reduced during culture, independent of the media used. In conclusion, under the conditions used in this study, the M199 and McCoy media are recommended for the culture of follicles derived from fresh and frozen ovarian tissues, respectively.

  12. Bovine meniscal tissue exhibits age- and interleukin-1 dose-dependent degradation patterns and composition-function relationships.

    PubMed

    Ling, Carrie H-Y; Lai, Janice H; Wong, Ivan J; Levenston, Marc E

    2016-05-01

    Despite increasing evidence that meniscal degeneration is an early event in the development of knee osteoarthritis, relatively little is known regarding the sequence or functional implications of cytokine-induced meniscal degradation or how degradation varies with age. This study examined dose-dependent patterns of interleukin-1 (IL-1)-induced matrix degradation in explants from the radially middle regions of juvenile and adult bovine menisci. Tissue explants were cultured for 10 days in the presence of 0, 1.25, 5, or 20 ng/ml recombinant human IL-1α. Juvenile explants exhibited immediate and extensive sulfated glycosaminoglycan (sGAG) loss and subsequent collagen release beginning after 4-6 days, with relatively little IL-1 dose-dependence. Adult explants exhibited a more graded response to IL-1, with dose-dependent sGAG release and a lower fraction of sGAG released (but greater absolute release) than juvenile explants. In contrast to juvenile explants, adult explants exhibited minimal collagen release over the 10-day culture. Compressive and shear moduli reflected the changes in explant composition, with substantial decreases for both ages but a greater relative decrease in juvenile tissue. Dynamic moduli exhibited stronger dependence on explant sGAG content for juvenile tissue, likely reflecting concomitant changes to both proteoglycan and collagen tissue components. The patterns of tissue degradation suggest that, like in articular cartilage, meniscal proteoglycans may partially protect collagen from cell-mediated degeneration. A more detailed view of functional changes in meniscal tissue mechanics with degeneration will help to establish the relevance of in vitro culture models and will advance understanding of how meniscal degeneration contributes to overall joint changes in early stage osteoarthritis. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:801-811, 2016. © 2015 Orthopaedic Research Society. Published by

  13. Pericardium-6 Acupressure for the Prevention of Postoperative Nausea and Vomiting

    DTIC Science & Technology

    1999-10-01

    two groups. Key Words: Acupressure, Postoperative Nausea and Vomiting, Pericardium-six Meridian, Alternative Medicine , Traditional Chinese...western medicine has been unable to definitively explain why this phenomenon continues to occur. Acupressure at the pericardium-six meridian has been... Medicine vi PERICARDIUM-6 ACUPRESSURE FOR THE PREVENTION OF POSTOPERATIVE NAUSEA AND VOMITING by Robert Michael Woods THESIS

  14. Xanthosine administration does not affect the proportion of epithelial stem cells in bovine mammary tissue, but has a latent negative effect on cell proliferation

    SciTech Connect

    Rauner, Gat; Barash, Itamar

    2014-10-15

    The challenge in manipulating the proportion of somatic stem cells lies in having to override tissue homeostasis. Xanthosine infusion via the teat canal has been reported to augment the number of label-retaining cells in the mammary gland of 3-month-old bovine calves. To further delineate xanthosine's effect on defined stem cells in the mammary gland of heifers—which are candidates for increased prospective milk production following such manipulation—bovine mammary parenchymal tissue was transplanted and integrated into the cleared mammary fat pad of immunodeficient mice. Xanthosine administration for 14 days did not affect the number of label-retaining cells after 10- and 11-week chases. No change in stem cell proportion, analyzed according to CD49f and CD24 expression, was noted. Clone formation and propagation rate of cultured cells, as well as expression of stem cell markers, were also unaffected. In contrast, a latent 50% decrease in bovine mammary cell proliferation rate was observed 11 weeks after xanthosine administration. Tumor development in mice was also limited by xanthosine administration. These effects may have resulted from an initial decrease in expression of the rate-limiting enzyme in guanine synthesis, IMPDH. The data indicate that caution should be exerted when considering xanthosine for stem cell manipulation. - Highlights: • Novel “bovinized“ mouse model for exogenous effects on bovine mammary gland. • Xanthosine did not affect stem cell number/function in bovine mammary gland. • Xanthosine caused an immediate decrease in IMPDH expression in bovine mammary gland. • Xanthosine had latent negative effect on cell proliferation in bovine mammary gland. • Xanthosine administration limited mammary tumor growth.

  15. A rare chronic constrictive pericarditis with localized adherent visceral pericardium and normal parietal pericardium: a case report.

    PubMed

    Ni, Qingqiang; Yun, Lin; Xu, Rui; Li, Guohua; Yao, Yucai; Li, Jiamin

    2016-09-01

    Classic constrictive pericarditis (CP) is characterized by fibrous scarring and adhesion of both the visceral pericardium and the parietal pericardium, which leads to restricted cardiac filling. However, diagnosing CP with normal thickness pericardium and without calcification is still a challenge. The predominant cause in the developed world is idiopathic or viral pericarditis followed by post-cardiac surgery and post-radiation. Tuberculosis still remains a common cause of CP in developing countries. In this report, we describe a rare case of idiopathic localized constrictive visceral pericardium with normal thickness of the parietal pericardium in a middle-aged man. The patient presented with unexplained right heart failure and echocardiography showed moderate bi-atrial enlargement which should be identified with the restrictive cardiomyopathy. After 10 months of conservative treatment, the progression of right heart failure was remaining. A pericardiectomy was performed and the patient recovered. This case serves as a reminder to consider CP in patients with unexplained right heart failure, so that timely investigation and treatment can be initiated.

  16. Hyaline cartilage tissue is formed through the co-culture of passaged human chondrocytes and primary bovine chondrocytes.

    PubMed

    Taylor, Drew W; Ahmed, Nazish; Hayes, Anthony J; Ferguson, Peter; Gross, Allan E; Caterson, Bruce; Kandel, Rita A

    2012-08-01

    To circumvent the problem of a sufficient number of cells for cartilage engineering, the authors previously developed a two-stage culture system to redifferentiate monolayer culture-expanded dedifferentiated human articular chondrocytes by co-culture with primary bovine chondrocytes (bP0). The aim of this study was to analyze the composition of the cartilage tissue formed in stage 1 and compare it with bP0 grown alone to determine the optimal length of the co-culture stage of the system. Biochemical data show that extracellular matrix accumulation was evident after 2 weeks of co-culture, which was 1 week behind the bP0 control culture. By 3 to 4 weeks, the amounts of accumulated proteoglycans and collagens were comparable. Expression of chondrogenic genes, Sox 9, aggrecan, and collagen type II, was also at similar levels by week 3 of culture. Immunohistochemical staining of both co-culture and control tissues showed accumulation of type II collagen, aggrecan, biglycan, decorin, and chondroitin sulfate in appropriate zonal distributions. These data indicate that co-cultured cells form cartilaginous tissue that starts to resemble that formed by bP0 after 3 weeks, suggesting that the optimal time to terminate the co-culture stage, isolate the now redifferentiated cells, and start stage 2 is just after 3 weeks.

  17. Binding of bovine thyrotropin to specific sites in thyroid tissue from control and hemithyroidectomized rats

    SciTech Connect

    Clark, O.H.; Lambert, W.R.; Amir, S.M.; Ingbar, S.H.

    1985-12-01

    The binding of 125I-bovine thyrotropin to thyroid particulate fractions from sham-operated (control) and hemithyroidectomized rats was compared to determine if a change in either the number of bovine thyroid-stimulating hormone (bTSH) binding sites or their affinity for bTSH occurs in physiological situations that evoke changes in the intensity of thyroid stimulation. Following hemithyroidectomy serum TSH levels increase and the remnant thyroid lobe enlarges. Because of compensatory thyroid hypertrophy the concentration of TSH binding sites in the thyroid glands from hemithyroidectomized and control rats was related to particulate protein concentration, to the degree of thyroid cellularity as indicated by DNA concentration, and to the concentration of the plasma membrane markers, 5'-nucleotidase and magnesium-dependent ATPase. In each of four experiments, saturation studies revealed that the maximum specific binding of TSH per unit particulate protein and per thyroid lobe was greater in particulates from remnant than from control thyroid lobes. When related to DNA concentration, the concentration of TSH binding sites in remnant lobes was approximately twice that in control lobes. Because of an increase in plasma membrane markers per lobe after hemithyroidectomy, however, there was no difference in the number of TSH binding sites when related to the concentrations of the membrane marker enzymes in the particulate fractions. As judged from Scatchard analysis, the affinity of TSH binding was lower in remnant than in control lobes. This was partially but not completely due to the increased concentration of particulate protein in the remnant thyroid. These experiments demonstrate that the increase in serum TSH levels after hemithyroidectomy in the rat is associated with alterations in TSH receptor capacity and affinity.

  18. Hypertrophy, hyperplasia, and infectious virus in gut-associated lymphoid tissue of mice after oral inoculation with simian-human or bovine-human reassortant rotaviruses.

    PubMed

    Moser, C A; Dolfi, D V; Di Vietro, M L; Heaton, P A; Offit, P A; Clark, H F

    2001-04-01

    Oral inoculation of infants with a vaccine that contains simian-human reassortant rotaviruses has been found to be a rare cause of intussusception. Because intussusception can be associated with enlargement of gut-associated lymphoid tissue, we studied the capacity of simian-human and bovine-human reassortant rotaviruses to cause lymphoid hypertrophy and hyperplasia of Peyer's patches (PP) of adult BALB/c mice. Neither hypertrophy nor hyperplasia was detected in PP after oral inoculation with simian-human or bovine-human reassortant rotaviruses. However, infectious virus was detected in PP and mesenteric lymph nodes after oral inoculation with simian, but not bovine, reassortant rotaviruses. Implications of these findings on the pathogenesis of intussusception are discussed.

  19. Polyamine synthesis in mammalian tissues. Isolation and characterization of spermidine synthase from bovine brain.

    PubMed Central

    Raina, A; Hyvönen, T; Eloranta, T; Voutilainen, M; Samejima, K; Yamanoha, B

    1984-01-01

    Spermidine synthase (EC 2.5.1.16) was purified to apparent homogeneity (about 11 000-fold) from bovine brain by affinity chromatography, with S-adenosyl-(5')-3-thiopropylamine linked to Sepharose as the adsorbent. The enzyme preparation was free from S-adenosylmethionine decarboxylase (EC 4.1.1.50) and spermine synthase (EC 2.5.1.22) activities. The native enzyme had an apparent Mr of 70 000, was composed of two subunits of equal size, and had an isoelectric point at pH 5.22. The apparent Km values for putrescine and decarboxylated adenosylmethionine [S-adenosyl-(5')-3-methylthiopropylamine] were 40 microM and 0.3 microM respectively. Cadaverine and 1,6-diaminohexane could replace putrescine as the aminopropyl acceptor, although the reaction rates were only 6% and 1% respectively of that obtained with putrescine. Ethyl, propyl and carboxymethyl analogues of decarboxy-S-adenosylmethionine could act as propylamine donors. Both the reaction products, spermidine and 5'-methylthioadenosine, were mixed-type inhibitors of the enzyme. On the basis of initial-velocity and product-inhibition studies, a ping-pong reaction mechanism for the spermidine synthase reaction was ruled out. Images Fig. 1. PMID:6743257

  20. Production of Bovine Embryos and Calves Cloned by Nuclear Transfer Using Mesenchymal Stem Cells from Amniotic Fluid and Adipose Tissue.

    PubMed

    da Silva, Carolina Gonzales; Martins, Carlos Frederico; Cardoso, Tereza Cristina; da Cunha, Elisa Ribeiro; Bessler, Heidi Christina; Martins, George Henrique Lima; Pivato, Ivo; Báo, Sônia Nair

    2016-04-01

    The less differentiated the donor cells are used in nuclear transfer (NT), the more easily are they reprogrammed by the recipient cytoplasm. In this context, mesenchymal stem cells (MSCs) appear as an alternative to donor nuclei for NT. The amniotic fluid and adipose tissue are sources of MSCs that have not been tested for the production of cloned embryos in cattle. The objective of this study was to isolate, characterize, and use MSCs derived from amniotic fluid (MSC-AF) and adipose tissue (MSC-AT) to produce cloned calves. Isolation of MSC-AF was performed using in vivo ultrasound-guided transvaginal amniocentesis, and MSC-AT were isolated by explant culture. Cellular phenotypic and genotypic characterization by flow cytometry, immunohistochemistry, and RT-PCR were performed, as well as induction in different cell lineages. The NT was performed using MSC-AF and MSC-AT as nuclear donors. The mesenchymal markers of MSC were expressed in bovine MSC-AF and MSC-AT cultures, as evidenced by flow cytometry, immunohistochemistry, and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes, and adipocytes. Embryo production was similar between the cell types, and two calves were born. The calf from MSC-AT was born healthy, and this fact opens a new possibility of using this type of cell to produce cloned cattle by NT.

  1. Endometrial cells sense and react to tissue damage during infection of the bovine endometrium via interleukin 1

    PubMed Central

    Healy, Laura L.; Cronin, James G.; Sheldon, I. Martin

    2014-01-01

    Cells generate inflammatory responses to bacteria when pattern recognition receptors bind pathogen-associated molecules such as lipopolysaccharide. Cells may also respond to tissue damage by sensing damage-associated molecules. Postpartum bacterial infections of the bovine uterus cause endometritis but the risk of disease is increased by tissue trauma triggered by dystocia. Animals that suffered dystocia had increased concentrations of inflammatory mediators IL-8, IL-1β and IL-1α in vaginal mucus 3 weeks postpartum, but they also had more bacteria than normal animals. Ex vivo organ cultures of endometrium, endometrial cells and peripheral blood monocytes did not generate inflammatory responses to prototypical damage molecules, HMGB1 or hyaluronan, or to necrotic cells; although they secreted IL-6 and IL-8 in a concentration-dependent manner when treated with IL-1α. However, necrotic endometrial cells did not accumulate intracellular IL-1α or release IL-1α, except when pre-treated with lipopolysaccharide or bacteria. Endometrial cell inflammatory responses to IL-1α were dependent on the cognate receptor IL-1R1, and the receptor adaptor protein MyD88, and the inflammatory response to IL-1α was independent of the response to lipopolysaccharide. Rather than a typical damage-associated molecule, IL-1α acts to scale the inflammatory response in recognition that there is a combination of pathogen challenge followed by endometrial cell damage. PMID:25395028

  2. Endometrial cells sense and react to tissue damage during infection of the bovine endometrium via interleukin 1.

    PubMed

    Healy, Laura L; Cronin, James G; Sheldon, I Martin

    2014-11-14

    Cells generate inflammatory responses to bacteria when pattern recognition receptors bind pathogen-associated molecules such as lipopolysaccharide. Cells may also respond to tissue damage by sensing damage-associated molecules. Postpartum bacterial infections of the bovine uterus cause endometritis but the risk of disease is increased by tissue trauma triggered by dystocia. Animals that suffered dystocia had increased concentrations of inflammatory mediators IL-8, IL-1β and IL-1α in vaginal mucus 3 weeks postpartum, but they also had more bacteria than normal animals. Ex vivo organ cultures of endometrium, endometrial cells and peripheral blood monocytes did not generate inflammatory responses to prototypical damage molecules, HMGB1 or hyaluronan, or to necrotic cells; although they secreted IL-6 and IL-8 in a concentration-dependent manner when treated with IL-1α. However, necrotic endometrial cells did not accumulate intracellular IL-1α or release IL-1α, except when pre-treated with lipopolysaccharide or bacteria. Endometrial cell inflammatory responses to IL-1α were dependent on the cognate receptor IL-1R1, and the receptor adaptor protein MyD88, and the inflammatory response to IL-1α was independent of the response to lipopolysaccharide. Rather than a typical damage-associated molecule, IL-1α acts to scale the inflammatory response in recognition that there is a combination of pathogen challenge followed by endometrial cell damage.

  3. Determination of sulfamethazine in bovine and porcine tissues by reversed-phase liquid chromatography.

    PubMed

    Boison, J O; Keng, L J

    1994-01-01

    A simple, sensitive, and rapid method for the liquid chromatographic determination of sulfamethazine in animal tissues was developed by using sulfaethoxypyridazine as the internal standard. Homogenized tissue is extracted with chloroform, and the sulfa drugs are back-extracted from chloroform into alkaline sodium chloride solution. The pH of the aqueous extract is adjusted to 6, and the sulfas are concentrated on a conditioned C18 cartridge and eluted with 1 mL methanol. Sulfamethazine and sulfaethoxypyridazine are separated from tissue co-extractives by reversed-phase chromatography on a C18 column by using 0.05M sodium dihydrogen phosphate-methanol (7 + 3). Detection is performed at 265 nm. The method has a detection limit of 2 ng/g. Results obtained by this method were compared with those obtained by the official thin-layer chromatography/densitometric method.

  4. Influence of ultrasonic scattering in the calculation of thermal dose in ex-vivo bovine muscular tissues.

    PubMed

    Cortela, Guillermo A; von Krüger, Marco A; Negreira, Carlos A; Pereira, Wagner C A

    2016-02-01

    This study explores the effect of ultrasound scattering on the temperature increase in phantoms and in samples of ex-vivo biological tissue through the calculation of the thermal dose (TD). Phantoms with different weight percentages of graphite powder (0-1%w/w, different scattering mean free paths, ℓS) and ex-vivo bovine muscle tissue were isonified by therapeutic ultrasound (1 MHz). The TD values were calculated from the first 4 min of experimental temperature curves obtained at several depths and were compared with those acquired from the numerical solution of the bio-heat transfer equation (simulated with 1 MHz and 0.5-2.0 W cm(-2)). The temperature curves suggested that scattering had an important role because the temperature increments were found to be higher for higher percentages of graphite powder (lower ℓS). For example, at a 30-mm depth and a 4-min therapeutic ultrasound application (0.5 W cm(-2)), the TDs (in equivalent minutes at 43 °C) were 7.2, 17.8, and 58.3 for the phantom with ℓS of 4.35, 3.85, and 3.03 mm, respectively. In tissue, the inclusion of only absorption or full attenuation in the bio-heat transfer equation (BHTE) heat source term of the simulation leads to under- or overestimation of the TD, respectively, as compared to the TD calculated from experimental data. The experiments with phantoms (with different scatterer concentrations) and ex-vivo samples show that the high values of TD were caused by the increase of energy absorption due to the lengthening of the propagation path caused by the changing in the propagation regime.

  5. Isolation, in vitro culture and identification of a new type of mesenchymal stem cell derived from fetal bovine lung tissues.

    PubMed

    Hu, Pengfei; Pu, Yabin; Li, Xiayun; Zhu, Zhiqiang; Zhao, Yuhua; Guan, Weijun; Ma, Yuehui

    2015-09-01

    Lung‑derived mesenchymal stem cells (LMSCs) are considered to be important in lung tissue repair and regenerative processes. However, the biological characteristics and differentiation potential of LMSCs remain to be elucidated. In the present study, fetal lung‑derived mesenchymal stem cells (FLMSCs) were isolated from fetal bovine lung tissues by collagenase digestion. The in vitro culture conditions were optimized and stabilized and the self‑renewal ability and differentiation potential were evaluated. The results demonstrated that the FLMSCs were morphologically consistent with fibroblasts, were able to be cultured and passaged for at least 33 passages and the cell morphology and proliferative ability were stable during the first 10 passages. In addition, FLMSCs were found to express CD29, CD44, CD73 and CD166, however, they did not express hematopoietic cell specific markers, including CD34, CD45 and BOLA‑DRα. The growth kinetics of FLMSCs consisted of a lag phase, a logarithmic phase and a plateau phase, and as the passages increased, the proliferative ability of cells gradually decreased. The majority of FLMSCs were in G0/G1 phase. Following osteogenic induction, FLMSCs were positive for the expression of osteopontin and collagen type I α2. Following neurogenic differentiation, the cells were morphologically consistent with neuronal cells and positive for microtubule‑associated protein 2 and nestin expression. It was concluded that the isolated FLMSCs exhibited typical characteristics of mesenchymal stem cells and that the culture conditions were suitable for their proliferation and the maintenance of stemness. The present study illustrated the potential application of lung tissue as an adult stem cell source for regenerative therapies.

  6. Determination of ceftiofur metabolite desfuroylceftiofur cysteine disulfide in bovine tissues using liquid chromatography-tandem mass spectrometry as a surrogate marker residue for ceftiofur.

    PubMed

    Feng, Shixia; Chiesa, Oscar A; Kijak, Philip; Chattopadhaya, Chaitali; Lancaster, Vicki; Smith, Elizabeth A; Girard, Lauren; Sklenka, Sara; Li, Hui

    2014-06-04

    Ceftiofur is a widely used cephalosporin β-lactam antibiotic with frequently reported residue violations. This paper reports a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining a ceftiofur metabolite, desfuroylceftiofur cysteine disulfide (DCCD), in bovine kidney, liver, and muscle tissues. Incurred tissue samples were obtained from dosed animals and analyzed to evaluate the utility of the method. For kidney, the target tissue, the method utilized a simple extraction with phosphate buffer followed by solid phase extraction (SPE) cleanup. For liver and muscle, acetonitrile and hexane were used to remove most proteins and fat from the initial buffer extract before the SPE cleanup. Method accuracy was between 97 and 107%, and the coefficient of variation was between 3.4 and 11.0% for all three types of tissues. The relationship between the new and regulatory methods for bovine kidney was established. It was concluded that DCCD is a suitable surrogate marker residue for ceftiofur in bovine kidney.

  7. Sequence evaluation of four pooled-tissue normalized bovine cDNA libraries and construction of a gene index for cattle.

    PubMed

    Smith, T P; Grosse, W M; Freking, B A; Roberts, A J; Stone, R T; Casas, E; Wray, J E; White, J; Cho, J; Fahrenkrug, S C; Bennett, G L; Heaton, M P; Laegreid, W W; Rohrer, G A; Chitko-McKown, C G; Pertea, G; Holt, I; Karamycheva, S; Liang, F; Quackenbush, J; Keele, J W

    2001-04-01

    An essential component of functional genomics studies is the sequence of DNA expressed in tissues of interest. To provide a resource of bovine-specific expressed sequence data and facilitate this powerful approach in cattle research, four normalized cDNA libraries were produced and arrayed for high-throughput sequencing. The libraries were made with RNA pooled from multiple tissues to increase efficiency of normalization and maximize the number of independent genes for which sequence data were obtained. Target tissues included those with highest likelihood to have impact on production parameters of animal health, growth, reproductive efficiency, and carcass merit. Success of normalization and inter- and intralibrary redundancy were assessed by collecting 6000-23,000 sequences from each of the libraries (68,520 total sequences deposited in GenBank). Sequence comparison and assembly of these sequences was performed in combination with 56,500 other bovine EST sequences present in the GenBank dbEST database to construct a cattle Gene Index (available from The Institute for Genomic Research at http://www.tigr.org/tdb/tgi.shtml). The 124,381 bovine ESTs present in GenBank at the time of the analysis form 16,740 assemblies that are listed and annotated on the Web site. Analysis of individual library sequence data indicates that the pooled-tissue approach was highly effective in preparing libraries for efficient deep sequencing.

  8. Omega-6 Fat Supplementation Alters Lipogenic Gene Expression in Bovine Subcutaneous Adipose Tissue

    PubMed Central

    Joseph, Sandeep J.; Pratt, Scott L.; Pavan, Enrique; Rekaya, Romdhane; Duckett., Susan K.

    2010-01-01

    In contrast to rodents, adipose tissue serves as the major site of lipogenesis and storage reservoir for excess dietary energy in cattle. Research in rodents shows that adding corn oil (57% C18:2 n-6) to the diet alters lipogenesis enhancing deposition of omega-6 fatty acids. This study examines changes in lipogenic gene expression of subcutaneous adipose tissue from eighteen steers fed increasing levels of dietary corn oil [0 (NONE), 0.31 kg/d (MED) and 0.62 kg/d (HI)] using two platforms, qPCR and microarray. The results show that MED level of oil supplementation up-regulates gene expression of key lipogenic enzymes but that as oil supplementation reaches HI level mRNA encoding lipogenic enzymes responsible for de novo synthesis and desaturation are down-regulated. Changes in specific lipogenic mRNA levels are correlated with changes in tissue fatty acid composition where de novo and desatured fatty acids were reduced with the highest level of oil supplementation. PMID:21072324

  9. Fish meal supplementation increases bovine plasma and luteal tissue omega-3 fatty acid composition.

    PubMed

    White, N R; Burns, P D; Cheatham, R D; Romero, R M; Nozykowski, J P; Bruemmer, J E; Engle, T E

    2012-03-01

    The objective of this experiment was to determine if dietary inclusion of fish meal would increase plasma and luteal tissue concentrations of eicosapentaenoic and docosahexaenoic acids. Seventeen nonlactating Angus cows (2 to 8 yr of age) were housed in individual pens and fed a corn silage-based diet for approximately 60 d. Diets were supplemented with fish meal at 5% DMI (a rich source of eicosapentaenoic acid and docosahexaenoic acid; n = 9 cows) or corn gluten meal at 6% DMI (n = 8 cows). Body weights and jugular blood samples were collected immediately before the initiation of supplementation and every 7 d thereafter for 56 d to monitor plasma n-3 fatty acid composition and BW. Estrous cycles were synchronized using 2 injections of PGF(2α) administered at 14-d intervals. The ovary bearing the corpus luteum was surgically removed at midcycle (between d 10 and 12) after estrus synchronization, which corresponded to approximately d 60 of supplementation. The ovary was transported to the laboratory, and approximately 1.5 g of luteal tissue was stored at -80°C until analyzed for n-3 fatty acid content. Initial and ending BW did not differ (P > 0.10) between cows supplemented with fish meal and those with corn gluten meal. Plasma eicosapentaenoic acid was greater (P < 0.05) beginning at d 7 of supplementation and docosahexaenoic was greater (P < 0.05) beginning at d 14 of supplementation for cows receiving fish meal. Luteal tissue collected from fish meal-supplemented cows had greater (P < 0.05) luteal n-3 fatty acids and reduced (P < 0.05) arachidonic acid and n-6 to n-3 ratio as compared with tissue obtained from cows supplemented with corn gluten meal. Our data show that fish meal supplementation increases luteal n-3 fatty acid content and reduces available arachidonic acid content, the precursor for PGF(2α). The increase in luteal n-3 fatty acids may reduce PGF(2α) intraluteal synthesis after breeding resulting in increased fertility in cattle.

  10. A review of the structure of human and bovine dental hard tissues and their physicochemical behaviour in relation to erosive challenge and remineralisation.

    PubMed

    Laurance-Young, P; Bozec, L; Gracia, L; Rees, G; Lippert, F; Lynch, R J M; Knowles, J C

    2011-04-01

    This review sets out to examine the suitability of bovine hard dental material in lieu of human material when investigating dental erosion, to review the evidence for the major factors popularly attributed to dental erosion: pH, pKa, acid type, erosion duration, temperature and stirring rate as well as examine the case for the use of fluoride in an anti-erosion capacity. Published works were selected using online search software ICI Web of Knowledge and Pubmed, with key terms such as "enamel", "erosion" and "bovine AND human" and cross referenced with relevant papers cited in the indices. The growing trend of dental erosion, coupled to legislative changes has precipitated a recent shortage of human enamel and dentine for experimental work. This in turn has resulted in the increasing use of cheap and readily available alternate supplies being sourced. This alternate supply principally originates from beef cattle under 20 months of age, under the assumption that bovine enamel and dentine will behave in a manner similar to human material. Recent experiments attempting to compare the physicochemical properties of these two species have shown that erosion is not simply a matter of bulk tissue loss resulting from acid exposure, but a multi-factorial event encompassing ever increasing and varied complexity of the inter-relationship between solvent and substrate. Accurate data from the published literature regarding the comparative properties of human and bovine hard dental tissue remains scarce but consensus appears to accept the continuing use of bovine enamel as a substitute for human enamel. This lack of comparative data is further hampered by the lack of an established, standardised protocol with which to evaluate the two species. In addition, much debate remains regarding the significant principal factors responsible for dental erosion and ways to minimise the pathological manifestation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. The phytanic acid content of the lipids of bovine tissues and milk.

    PubMed

    Lough, A K

    1977-01-01

    In three steers which were given grass silage for six months, the content of phytanic acid (i.e. 3,7,11,15-tetramethyl-hexadecanoic acid) in plasma lipid increased to about 8% of the total fatty acids, whereas after this time the proportion in the total fatty acids of liver and heart lipids was about 1%, and only 0.1% in those of kidney lipids; the acid was present in trace amounts in adipose-tissue triglycerides and was apparently absent from brain lipids. In eight lactating cows which were given grass silage for about 3 months, the content of phytanic acid in the total long chain fatty acids of milk and of plasma was 0.7% and 13%, respectively. In the plasma lipids of both steers and lactating cows, phytanic acid constituted a substantial proportion of the total fatty acids of the triglycerides and phospholipids; the acid was present in lowest proportion in the cholesteryl esters.

  12. Tissue temperatures attained in indocyanine-green infiltrated and noninfiltrated bovine eyelids using diode laser (805-nm) energy at 3, 5, and 7 watts

    NASA Astrophysics Data System (ADS)

    Kirkpatrick, John G.; Bartels, Kenneth E.; Dickey, D. Thomas; Powell, Ronald; Schafer, Steven A.; Zhou, Jin Fu; Wicksted, James P.

    1998-07-01

    Normal non-pigmented bovine eyelids in two Hereford cows under general anesthesia were treated with diode laser (805 nm) in a power range of 3 - 6.5 watts. Tissue temperatures were measured in areas infiltrated with 0.25% indocyanine green (ICG) solution and in non-infiltrated areas. Targeted tissue was laser treated at post-injection time intervals of 1 to 60 minutes. Temperatures were measured with a computerized temperature sensing program using hypodermic needle thermistors. Trial objectives were to establish power/chromophore concentration/time parameters to create tumor and peri-tumor tissue temperatures of 50 degrees Celsius without causing epidermal vaporization. Tissue temperature of 50 degrees Celsius has been established as the temperature producing coagulative necrosis of squamous cell carcinoma tumor tissue in 30 seconds. This temperature was produced in chromophore-enhanced tissue bit without repeatable confidence. Bovine ocular squamous cell carcinoma (BOSCC)-affected eyes and eyelids have been successfully treated with carbon dioxide focused and defocused laser with and without chromophore enhanced diode laser (805 nm) energy. The need for tumor staging and procedure standardization requires further investigation to determine laser power/chromophore concentration, and issue exposure times.

  13. Detection of bovine viral diarrhoea virus infected cattle--testing tissue samples derived from ear tagging using an Erns capture ELISA.

    PubMed

    Kuhne, S; Schroeder, C; Holmquist, G; Wolf, G; Horner, S; Brem, G; Ballagi, A

    2005-08-01

    A new diagnostic approach testing tissue samples derived from cattle ear tagging for bovine viral diarrhoea virus (BVDV) antigen in a commercially available antigen capture enzyme-linked immunosorbent assay (ACE) was developed. To validate this method, 99 positive and 469 negative samples were tested. With those samples the assay yielded a sensitivity of 100% and specificity of >or=99.6%. Serum and ear tissue samples from 11 persistently infected (PI) BVDV calves were tested. While serum samples were negative after intake of colostrum, the ear tissue samples could be detected positive for BVDV all the time. Testing multiple samples derived from the same ear from PI cattle yielded positive results and low variation. Using cattle ear tags combining the ear tag application with sampling of a small ear tissue plug and testing those tissue samples with an ACE could be a reliable and economic way of BVDV testing.

  14. Aortic Arch Reconstruction in Neonates with Biventricular Morphology: Increased Risk for Development of Recoarctation by Use of Autologous Pericardium.

    PubMed

    Bechtold, Caroline; Purbojo, Ariawan; Schwitulla, Judith; Glöckler, Martin; Toka, Okan; Dittrich, Sven; Cesnjevar, Robert Anton; Rüffer, André

    2015-08-01

    The aim of this study was to analyze risk factors promoting development of recoarctation (Re-CoA) in neonates who survived aortic arch repair from an anterior approach. Fifty consecutive neonates with biventricular morphology and ductal-dependent lower body perfusion who were discharged home following aortic arch repair with cardiopulmonary bypass between 2000 and 2012 were retrospectively reviewed. Arch anatomy was either interruption (n = 10) or hypoplasia with coarctation (n = 40). Aortic arch reconstruction was performed by using patch material (bovine pericardium, n = 30, homograft, n = 10, or glutaraldehyde-treated autologous pericardium, n = 7), and three patients underwent direct end-to-side anastomosis. Antegrade cerebral and continuous myocardial perfusion was performed in 39 and 21 patients, respectively. Kaplan-Meier freedom from Re-CoA was calculated. Morphologic and perioperative data indicating increased risk of Re-CoA by univariate analysis were included in multivariate Cox regression analysis. Mean follow-up was 5.3 ± 4.1 years. Re-CoA occurred in 13 patients and was treated successfully by balloon dilatation (n = 6) or surgery (n = 7). Freedom from Re-CoA after 1 and 5 years was 83 ± 5 and 79 ± 6%, respectively. Two patients died early after surgical repair of Re-CoA. The use of autologous pericardium for aortic arch augmentation was the only independent risk factor for development of Re-CoA (hazard ratio: 4.3 [95% confidence interval: 1.2-16.1]; p = 0.028). Re-CoA following neonatal aortic arch surgery can be treated by balloon dilatation or surgery, if adequate. In this study, the risk for development of Re-CoA was independently increased by the use of autologous pericardium during initial arch repair. Georg Thieme Verlag KG Stuttgart · New York.

  15. Congenital absence of the pericardium presenting as acute myocardial necrosis

    PubMed Central

    Brulotte, Steeve; Roy, Louis; Larose, Eric

    2007-01-01

    Congenital absence of the pericardium is a rare defect of which recognition is critical because it can be associated with catastrophic outcomes. While some carriers of this condition may present with a lethal complication, most are asymptomatic, and the defect is found incidentally. The case of a 49-year-old woman is described who presented with acute myocardial necrosis and absence of obstructive coronary artery disease, and in whom a complete left pericardial defect was found. An investigation was undertaken to determine the role of the defect in this patient’s presentation and to recommend the best possible therapy. Finally, a review of literature focusing on congenital absence of the pericardium, as well as a discussion of clinical presentation, imaging techniques and therapeutic options, is also presented. PMID:17876387

  16. Age dependence of dielectric properties of bovine brain and ocular tissues in the frequency range of 400 MHz to 18 GHz

    NASA Astrophysics Data System (ADS)

    Schmid, Gernot; Überbacher, Richard

    2005-10-01

    In order to identify possible age-dependent dielectric properties of brain and eye tissues in the frequency range of 400 MHz to 18 GHz, measurements on bovine grey and white matter as well as on cornea, lens (cortical) and the vitreous body were performed using a commercially available open-ended coaxial probe and a computer-controlled vector network analyser. Freshly excised tissues of 52 animals of two age groups (42 adult animals, i.e. 16-24 month old and 10 young animals, i.e. 4-6 month old calves) were examined within 8 min (brain tissue) and 15 min (eye tissue), respectively, of the animals' death. Tissue temperatures for the measurements were 32 ± 1 °C and 25 ± 1 °C for brain and eye tissues, respectively. Statistical analysis of the measured data revealed significant differences in the dielectric properties of white matter and cortical lens tissue between the adult and the young group. In the case of white matter the mean values of conductivity and permittivity of young tissue were 15%-22% and 12%-15%, respectively, higher compared to the adult tissue in the considered frequency range. Similarly, young cortical lens tissue was 25%-76% higher in conductivity and 27%-39% higher in permittivity than adult cortical lens tissue.

  17. Tissue oxygenation after exchange transfusion with ultrahigh-molecular-weight tense- and relaxed-state polymerized bovine hemoglobins.

    PubMed

    Cabrales, Pedro; Zhou, Yipin; Harris, David R; Palmer, Andre F

    2010-03-01

    Hemoglobin (Hb)-based O(2) carriers (HBOCs) constitute a class of therapeutic agents designed to correct the O(2) deficit under conditions of anemia and traumatic blood loss. The O(2) transport capacity of ultrahigh-molecular-weight bovine Hb polymers (PolybHb), polymerized in the tense (T) state and relaxed (R) state, were investigated in the hamster chamber window model using microvascular measurements to determine O(2) delivery during extreme anemia. The anemic state was induced by hemodilution with a plasma expander (70-kDa dextran). After an initial moderate hemodilution to 18% hematocrit, animals were randomly assigned to exchange transfusion groups based on the type of PolybHb solution used (namely, T-state PolybHb and R-state PolybHb groups). Measurements of systemic parameters, microvascular hemodynamics, capillary perfusion, and intravascular and tissue O(2) levels were performed at 11% hematocrit. Both PolybHbs were infused at 10 g/dl, and their viscosities were higher than nondiluted blood. Restitution of the O(2) carrying capacity with T-state PolybHb exhibited lower arterial pressure and higher functional capillary density compared with R-state PolybHb. Central arterial O(2) tensions increased significantly for R-state PolybHb compared with T-state PolybHb; conversely, microvascular O(2) tensions were higher for T-state PolybHb compared with R-state PolybHb. The increased tissue Po(2) attained with T-state PolybHb results from the larger amount of O(2) released from the PolybHb and maintenance of macrovascular and microvascular hemodynamics compared with R-state PolybHb. These results suggest that the extreme high O(2) affinity of R-state PolybHb prevented O(2) bound to PolybHb from been used by the tissues. The results presented here show that T-state PolybHb, a high-viscosity O(2) carrier, is a quintessential example of an appropriately engineered O(2) carrying solution, which preserves vascular mechanical stimuli (shear stress) lost during anemic

  18. A Simple and Effective Technique for Anatomical Approximation of the Upper Pericardium Using Hemostatic Clips.

    PubMed

    Husain, Mubassher; Acharya, Metesh Nalin; Cummings, Ian; Raja, Shahzad Gull

    Closure of the pericardium is important to protect bypass grafts, the great vessels, and the heart from injury due to sternal dehiscence. Furthermore, it is reported to reduce the formation of pericardial adhesions and thus facilitate entry into the chest at resternotomy. We here describe a simple, reproducible, and effective technique for tension-free approximation of the upper pericardium by applying small hemostatic clips to the preserved thymic fascia overlying the pericardium.

  19. Congenital Complete Absence of Pericardium Masquerading as Pulmonary Embolism

    PubMed Central

    Tariq, Saad; Mahmood, Sultan; Madeira, Samuel; Tarasov, Ethan

    2013-01-01

    Congenital absence of the pericardium is a rare cardiac condition, which can be either isolated or associated with other cardiac and extracardiac anomalies. There are six different types, depending on the severity of the involvement. Most of the patients with this defect are asymptomatic, especially the ones with complete absence of the pericardium. However, some patients are symptomatic, reporting symptoms that include chest pain, palpitations, dyspnea, and syncope. Diagnosis is established by the characteristic features on chest X-ray, echocardiogram, chest computed tomography (CT), and/or cardiac magnetic resonance imging (MRI). We present here a case of a 23 year-old-male, who presented to our hospital with complaints of pleuritic chest pain and exertional dyspnea, of a two-week duration. He was physically active and his past history was otherwise insignificant. His chest CT with contrast was interpreted as showing evidence of multiple emboli, predominantly in the left lung, and he was started on a heparin and warfarin therapy. A repeat chest CT with contrast three weeks later showed no significant change from the previous CT scan. Both scans showed that the heart was abnormally rotated to the left side of the chest. An echocardiogram raised the suspicion of congenital absence of the pericardium, with a posteriorly displaced heart. In retrospect, motion artifact on the left lung, attributed to cardiac pulsations and the lack of pericardium, resulted in a CT chest appearance, mimicking findings of pulmonary embolism. The misdiagnosis of pulmonary embolism was attributed to the artifact caused by excessive cardiac motion artifact on the chest CT scan. In non-gated CT angiograms, excessive motion causes an artifact that blurs the pulmonary vessels, reminiscent of a ′seagull′ or a ′boomerang′. Physicians need to be aware of this phenomenon, as well as the characteristic radiological features of this congenital anomaly, to enable them to make a correct

  20. Diet-induced changes in Ucp1 expression in bovine adipose tissues.

    PubMed

    Asano, Hiroki; Yamada, Tomoya; Hashimoto, Osamu; Umemoto, Takenao; Sato, Ryo; Ohwatari, Shiori; Kanamori, Yohei; Terachi, Tomohiro; Funaba, Masayuki; Matsui, Tohru

    2013-04-01

    Brown adipocytes, which regulate non-shivering thermogenesis, have been believed to exist in a limited number of mammalian species, and only under limited physiological conditions. Recent discoveries indicate that adult humans possess a significant number of functional brown adipocytes. This study explores the regulatory emergence of brown adipocytes in white adipose tissue (WAT) depots of fattening cattle. RT-PCR analyses indicated significant expression of Ucp1, a brown adipocyte-specific gene, in the WAT of 31-month-old Japanese Black steers. Immunohistochemical analysis revealed that Ucp1-positive small adipocytes were dispersed in the subcutaneous WAT. Next, we examined expression level of Ucp1 and other brown adipocyte-selective genes such as Pgc1α, Cidea, Dio2, Cox1, Cox7a1 and Cox8b in WAT of 30-month-old steers fed either diet with low protein/energy content (roughage diet) or that with high protein/energy content (concentrate diet) for 20months. Ucp1 expression in the subcutaneous WAT was significantly higher in the concentrate diet group than in the roughage diet group. Furthermore, the higher Ucp1 expression levels were limited to the subcutaneous WAT, and no differences between groups were detected in the mesenteric, perirenal, intermuscular or intramuscular WAT. Expression of Dio2, Cox1 and Cox8b was higher in the subcutaneous WAT but not in the mesenteric WAT of the concentrate diet group. Furthermore, expression of Prdm16, a positive regulator of differentiation toward brown adipocyte-lineage cells, and expression of leptin, a molecule that enhances activity of brown adipocytes, were significantly higher in the subcutaneous WAT of the concentrate diet group. This study demonstrates the presence of brown adipocytes in WAT depots of fattening cattle, and suggests the diet-related modulation of expression of genes predominantly expressed in brown adipocytes. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. Evaluation of tensile strength of tissue adhesives and sutures for clear corneal incisions using porcine and bovine eyes, with a novel standardized testing platform.

    PubMed

    Kaja, Simon; Goad, Daryl L; Ali, Fatima; Abraham, Ashley; Rebenitsch, R Luke; Teymoorian, Savak; Krishna, Rohit; Koulen, Peter

    2012-01-01

    Tissue adhesives for ophthalmologic applications were proposed almost 50 years ago, yet to date no adequate tissue glues have been identified that combine strong sealing properties with adequate safety and absence of postsurgical side effects. In recent years, cataract surgeries and Descemet's stripping with endothelial keratoplasty procedures have significantly increased the number of clear corneal incisions performed. One of the obstacles to discovery and development of novel tissue adhesives has been the result of nonstandardized testing of potential tissue glues. We developed an instrument capable of controlling intraocular pressure in explanted porcine and bovine eyes in order to evaluate sealants, adhesives, and surgical closure methods used in ophthalmic surgery in a controlled, repeatable, and validated fashion. We herein developed and validated our instrument by testing the adhesive properties of cyanoacrylate glue in both porcine and bovine explant eyes. The instrument applied and maintained intraocular pressure through a broad range of physiological intraocular pressures. Cyanoacrylate-based glues showed significantly enhanced sealing properties of clear corneal incisions compared with sutured wounds. This study shows the feasibility of our instrument for reliable and standardized testing of tissue adhesive for ophthalmological surgery.

  2. Rapid analysis of aminoglycoside antibiotics in bovine tissues using disposable pipette extraction and ultrahigh performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Lehotay, Steven J; Mastovska, Katerina; Lightfield, Alan R; Nuñez, Alberto; Dutko, Terry; Ng, Chilton; Bluhm, Louis

    2013-10-25

    A high-throughput qualitative screening and identification method for 9 aminoglycosides of regulatory interest has been developed, validated, and implemented for bovine kidney, liver, and muscle tissues. The method involves extraction at previously validated conditions, cleanup using disposable pipette extraction, and analysis by a 3 min ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. The drug analytes include neomycin, streptomycin, dihydrosptreptomycin, and spectinomycin, which have residue tolerances in bovine in the US, and kanamicin, gentamicin, apramycin, amikacin, and hygromycin, which do not have US tolerances established in bovine tissues. Tobramycin was used as an internal standard. An additional drug, paromomycin also was validated in the method, but it was dropped during implementation due to conversion of neomycin into paromomycin. Proposed fragmentation patterns for the monitored ions of each analyte were elucidated with the aid of high resolution MS using a quadrupole-time-of-flight instrument. Recoveries from spiking experiments at regulatory levels of concern showed that all analytes averaged 70-120% recoveries in all tissues, except hygromycin averaged 61% recovery. Lowest calibrated levels were as low as 0.005 μg/g in matrix extracts, which approximately corresponded to the limit of detection for screening purposes. Drug identifications at levels <0.05 μg/g were made in spiked and/or real samples for all analytes and tissues tested. Analyses of 60 samples from 20 slaughtered cattle previously screened positive for aminoglycosides showed that this method worked well in practice. The UHPLC-MS/MS method has several advantages compared to the previous microbial inhibition screening assay, especially for distinguishing individual drugs from a mixture and improving identification of gentamicin in tissue samples.

  3. Effects of addition of tissue-type plasminogen activator in in vitro fertilization medium on bovine embryo development and quality.

    PubMed

    Krania, F; Dovolou, E; Rekkas, C A; Theodosiadou, E K; Pappas, I; Amiridis, G S

    2015-02-01

    Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue-type plasminogen activator (t-PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus-oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t-PA and/or its inhibitor epsilon-aminocaproic acid (control, t-PA, t-PA+ε-ACA, ε-ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t-PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε-ACA. PAA in the treated group was significantly reduced by ε-ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t-PA-modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t-PA. In conclusion, it appears that excessive t-PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.

  4. Genome-wide DNA Methylation Profiles and Their Relationships with mRNA and the microRNA Transcriptome in Bovine Muscle Tissue (Bos taurine)

    PubMed Central

    Huang, Yong-Zhen; Sun, Jia-Jie; Zhang, Liang-Zhi; Li, Cong-Jun; Womack, James E.; Li, Zhuan-Jian; Lan, Xian-Yong; Lei, Chu-Zhao; Zhang, Chun-Lei; Zhao, Xin; Chen, Hong

    2014-01-01

    DNA methylation is a key epigenetic modification in mammals and plays important roles in muscle development. We sampled longissimus dorsi muscle (LDM) from a well-known elite native breed of Chinese Qinchuan cattle living within the same environment but displaying distinct skeletal muscle at the fetal and adult stages. We generated and provided a genome-wide landscape of DNA methylomes and their relationship with mRNA and miRNA for fetal and adult muscle studies. Integration analysis revealed a total of 77 and 1,054 negatively correlated genes with methylation in the promoter and gene body regions, respectively, in both the fetal and adult bovine libraries. Furthermore, we identified expression patterns of high-read genes that exhibit a negative correlation between methylation and expression from nine different tissues at multiple developmental stages of bovine muscle-related tissue or organs. In addition, we validated the MeDIP-Seq results by bisulfite sequencing PCR (BSP) in some of the differentially methylated promoters. Together, these results provide valuable data for future biomedical research and genomic and epigenomic studies of bovine skeletal muscle that may help uncover the molecular basis underlying economically valuable traits in cattle. This comprehensive map also provides a solid basis for exploring the epigenetic mechanisms of muscle growth and development. PMID:25306978

  5. Xanthosine administration does not affect the proportion of epithelial stem cells in bovine mammary tissue, but has a latent negative effect on cell proliferation.

    PubMed

    Rauner, Gat; Barash, Itamar

    2014-10-15

    The challenge in manipulating the proportion of somatic stem cells lies in having to override tissue homeostasis. Xanthosine infusion via the teat canal has been reported to augment the number of label-retaining cells in the mammary gland of 3-month-old bovine calves. To further delineate xanthosine׳s effect on defined stem cells in the mammary gland of heifers-which are candidates for increased prospective milk production following such manipulation-bovine mammary parenchymal tissue was transplanted and integrated into the cleared mammary fat pad of immunodeficient mice. Xanthosine administration for 14 days did not affect the number of label-retaining cells after 10- and 11-week chases. No change in stem cell proportion, analyzed according to CD49f and CD24 expression, was noted. Clone formation and propagation rate of cultured cells, as well as expression of stem cell markers, were also unaffected. In contrast, a latent 50% decrease in bovine mammary cell proliferation rate was observed 11 weeks after xanthosine administration. Tumor development in mice was also limited by xanthosine administration. These effects may have resulted from an initial decrease in expression of the rate-limiting enzyme in guanine synthesis, IMPDH. The data indicate that caution should be exerted when considering xanthosine for stem cell manipulation. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Genome-wide DNA methylation profiles and their relationships with mRNA and the microRNA transcriptome in bovine muscle tissue (Bos taurine).

    PubMed

    Huang, Yong-Zhen; Sun, Jia-Jie; Zhang, Liang-Zhi; Li, Cong-Jun; Womack, James E; Li, Zhuan-Jian; Lan, Xian-Yong; Lei, Chu-Zhao; Zhang, Chun-Lei; Zhao, Xin; Chen, Hong

    2014-10-13

    DNA methylation is a key epigenetic modification in mammals and plays important roles in muscle development. We sampled longissimus dorsi muscle (LDM) from a well-known elite native breed of Chinese Qinchuan cattle living within the same environment but displaying distinct skeletal muscle at the fetal and adult stages. We generated and provided a genome-wide landscape of DNA methylomes and their relationship with mRNA and miRNA for fetal and adult muscle studies. Integration analysis revealed a total of 77 and 1,054 negatively correlated genes with methylation in the promoter and gene body regions, respectively, in both the fetal and adult bovine libraries. Furthermore, we identified expression patterns of high-read genes that exhibit a negative correlation between methylation and expression from nine different tissues at multiple developmental stages of bovine muscle-related tissue or organs. In addition, we validated the MeDIP-Seq results by bisulfite sequencing PCR (BSP) in some of the differentially methylated promoters. Together, these results provide valuable data for future biomedical research and genomic and epigenomic studies of bovine skeletal muscle that may help uncover the molecular basis underlying economically valuable traits in cattle. This comprehensive map also provides a solid basis for exploring the epigenetic mechanisms of muscle growth and development.

  7. Morphometrical analysis of preantral follicular survival of VEGF-treated bovine ovarian cortex tissue following xenotransplantation in an immune deficient mouse model.

    PubMed

    Langbeen, A; Van Ginneken, C; Fransen, E; Bosmans, E; Leroy, J L M R; Bols, P E J

    2016-05-01

    The increasing number of cancer survivors the past decades, has sparked the need for fertility preservation strategies. Due to predominantly ethical constraints, human research material is scarce. A bovine in vitro model is a valuable alternative. Therefore, the following objectives were defined: 1) to xeno-graft bovine ovarian cortex tissue in immune deficient mice as a study-model for female fertility preservation strategies; 2) to stereologically quantify vascularization in Vascular Endothelial Growth Factor (VEGF)-treated and non-treated tissue; 3) to study preantral follicular survival in situ, after xenotransplantation. Bovine ovarian tissue strips were incubated with or without VEGF prior to grafting into female, neutered BALB/c-nu mice (n=16). Non-transplanted cortical tissue was used as a control. At time zero (control), two (2 weeks) and four (4 weeks) weeks after transplantation, grafts were retrieved and assessed by von Willebrand Factor and caspase-3 immunostaining. Data were analyzed using a linear mixed model. In the VEGF+ grafts, 31% of the follicles were considered 'alive' 2 weeks after transplantation, compared to only 17% in the VEGF- grafts (P<0.05). However, no difference could be detected 4 weeks after transplantation (P=0.76) with less follicles being considered 'alive' after transplantation (22%), compared to the control (47.5%) (P<0.05). Finally, the vascular surface density was significantly less in the grafts, irrespective of the transplantation period or the use of VEGF. Although the transplantation process overall negatively influenced the number of viable follicles and vascular density, VEGF exposure prior to transplantation can favor follicle survival during a 2 weeks transplantation period.

  8. Development of an indirect immunofluorescence assay for diagnosis of bovine viral diarrhoea virus on ear notch tissue samples in cattle infected persistently.

    PubMed

    Bedeković, Tomislav; Lemo, Nina; Lojkić, Ivana; Cvetnić, Zeljko; Cač, Zeljko; Madić, Josip

    2011-12-01

    Bovine viral diarrhoea virus (BVDV) causes a disease that has a wide range of clinical symptoms in domestic and wild ruminants. It is a major problem in cattle and causes significant economic losses in the cattle industry. The virus infects bovines of all ages and causes both immunosuppression and reproductive, respiratory and digestive disorders. Cattle infected persistently, as a continuing source of the virus and the main factor in transmission of the disease between and among herds, are the main source of BVDV and a primary factor in the epidemiology of the disease. To determine whether a BVDV infection is persistent, two samples should be taken at 3-4 week intervals and tested for the virus antigen. Animal sera, whole blood, organ and ear notch tissue samples can be used for BVDV diagnosis. In ear notch tissue, viral antigen can be detected by an antigen enzyme-linked immunosorbent assay (antigen ELISA), immunohistochemistry (IHC) and reverse-transcription polymerase chain reaction (RT-PCR). This paper describes the development and implementation of an indirect immunofluorescence (IF) method using ear notch tissue samples for diagnosis of cattle infected persistently. Results obtained by this method show that IF is a good alternative to RT-PCR and antigen ELISA and can be a quick and accurate method in diagnosis of BVDV in cattle infected persistently with this virus.

  9. Fatigue behaviour of young ostrich pericardium.

    PubMed

    Claramunt, Rafael; García Páez, José María; Alvarez, Lourdes; Ros, Antonio; Casado, María Concepción

    2012-08-01

    Young ostrich pericardia (biomaterial under study for manufacturing cardiac valve leaflets), has been subjected to biaxial tension fatigue until breakage. Supraphysiological values of pressure (1 to 6 atm) have been employed to accelerate damage and, therefore, to reduce testing time but at physiological frequency in order to avoid viscoelastic behaviour changes. The lifetime fatigue curves have been obtained and large scatter has been observed in the results but this can be strongly reduced with adequate material selection. The thickness-based selection of samples has proved to be ineffective both in reducing scatter or improving strength, but the energy-based selection aided with statistical decision techniques has been shown to be very successful. The energy loss (energy under the hysteresis loop of each load and unload cycle) appears to be a very accurate predictor of the expected fatigue lifetime of the tissue. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Screening of Danofloxacin residue in bovine tissue by terbium-sensitized luminescence on C18 sorbent strips

    USDA-ARS?s Scientific Manuscript database

    Danofloxacin (DANO) residue in bovine muscle was screened at 200 ng/g by terbium-sensitized luminescence (TSL) directly measured on 10x6 mm C18 sorbent strips. The analyte was first adsorbed on sorbent surface by immersion in defatted homogenates. After reagent application and desiccation, TSL was d...

  11. Detection of PrP(Sc) in peripheral tissues of clinically affected cattle after oral challenge with bovine spongiform encephalopathy

    USDA-ARS?s Scientific Manuscript database

    Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative prion disease that affects cattle and can be transmitted to human beings as new variant Creutzfeldt-Jakob disease (vCJD). A protease-resistant, disease-associated isoform of the prion protein (PrP**Sc) accumulates in the central ner...

  12. Bovine latent transforming growth factor beta 1-binding protein 2: molecular cloning, identification of tissue isoforms, and immunolocalization to elastin-associated microfibrils.

    PubMed Central

    Gibson, M A; Hatzinikolas, G; Davis, E C; Baker, E; Sutherland, G R; Mecham, R P

    1995-01-01

    Monoclonal antibodies to fibrillin 1 (MP340), a component of elastin-associated microfibrils, were used to screen cDNA libraries made from bovine nuchal ligament mRNA. One of the selected clones (cL9; 1.2 kb) hybridized on Northern (RNA) blotting with nuchal ligament mRNA to two abundant mRNAs of 9.0 and 7.5 kb, which were clearly distinct from fibrillin mRNA (10 kb). Further library screening and later reverse transcription PCR by the rapid amplification of cDNA ends (RACE) technique resulted in the isolation of additional overlapping cDNAs corresponding to about 6.7 kb of the mRNA. The encoded protein exhibited sequence similarity of around 80% with a recently identified human protein named latent transforming growth factor beta 1 (TGF-beta 1)-binding protein 2 (LTBP-2), indicating that the new protein was bovine LTBP-2. This was confirmed by the specific localization of bovine LTBP-2 cDNA probes to human chromosome 14q24.3, which is the locus of the human LTBP-2 gene. The domain structure of bovine LTBP-2 is very similar to that of the human LTBP-2, containing 20 examples of 6-cysteine epidermal growth factor-like repeats, 16 of which have the consensus sequence for calcium binding, together with 4 examples of 8-cysteine motifs characteristic of fibrillins and LTBP-1. A 4-cysteine sequence which is unique to bovine LTBP-2 and which has similarity to the 8-cysteine motifs was also present. Antibodies raised to two unique bovine LTBP-2 peptides specifically localized in tissue sections to the elastin-associated microfibrils, indicating that LTBP-2 is closely associated with these structures. Immunoblotting experiments identified putative LTBP-2 isoforms as a 260-kDa species released into the medium by cultured elastic tissue cells and as larger 290- and 310-kDa species in tissue extracts. A major proportion of tissue-derived LTBP-2 required treatment with 6 M guanidine for solubilization, indicating that the protein was strongly bound to the microfibrils. Most of

  13. Bovine latent transforming growth factor beta 1-binding protein 2: molecular cloning, identification of tissue isoforms, and immunolocalization to elastin-associated microfibrils.

    PubMed

    Gibson, M A; Hatzinikolas, G; Davis, E C; Baker, E; Sutherland, G R; Mecham, R P

    1995-12-01

    Monoclonal antibodies to fibrillin 1 (MP340), a component of elastin-associated microfibrils, were used to screen cDNA libraries made from bovine nuchal ligament mRNA. One of the selected clones (cL9; 1.2 kb) hybridized on Northern (RNA) blotting with nuchal ligament mRNA to two abundant mRNAs of 9.0 and 7.5 kb, which were clearly distinct from fibrillin mRNA (10 kb). Further library screening and later reverse transcription PCR by the rapid amplification of cDNA ends (RACE) technique resulted in the isolation of additional overlapping cDNAs corresponding to about 6.7 kb of the mRNA. The encoded protein exhibited sequence similarity of around 80% with a recently identified human protein named latent transforming growth factor beta 1 (TGF-beta 1)-binding protein 2 (LTBP-2), indicating that the new protein was bovine LTBP-2. This was confirmed by the specific localization of bovine LTBP-2 cDNA probes to human chromosome 14q24.3, which is the locus of the human LTBP-2 gene. The domain structure of bovine LTBP-2 is very similar to that of the human LTBP-2, containing 20 examples of 6-cysteine epidermal growth factor-like repeats, 16 of which have the consensus sequence for calcium binding, together with 4 examples of 8-cysteine motifs characteristic of fibrillins and LTBP-1. A 4-cysteine sequence which is unique to bovine LTBP-2 and which has similarity to the 8-cysteine motifs was also present. Antibodies raised to two unique bovine LTBP-2 peptides specifically localized in tissue sections to the elastin-associated microfibrils, indicating that LTBP-2 is closely associated with these structures. Immunoblotting experiments identified putative LTBP-2 isoforms as a 260-kDa species released into the medium by cultured elastic tissue cells and as larger 290- and 310-kDa species in tissue extracts. A major proportion of tissue-derived LTBP-2 required treatment with 6 M guanidine for solubilization, indicating that the protein was strongly bound to the microfibrils. Most of

  14. Relationship between stearoyl-CoA desaturase 1 gene expression, relative protein abundance, and its fatty acid products in bovine tissues.

    PubMed

    Rezamand, Pedram; Watts, Jason S; Yavah, Katherine M; Mosley, Erin E; Ma, Liying; Corl, Benjamin A; McGuire, Mark A

    2014-08-01

    Stearoyl-CoA desaturase 1 (SCD1) greatly contributes to the unsaturated fatty acids present in milk and meat of cattle. The SCD1 enzyme introduces a double bond into certain saturated fatty acyl-CoAs producing monounsaturated fatty acids (MUFA). The SCD1 enzyme also has been shown to be active in the bovine mammary gland converting t11 18:1 (vaccenic acid) to c9 t11 conjugated linoleic acid (CLA). The objective of this study was to determine any association between the gene expression of SCD1 and occurrence of its products (c9 14:1, c9 16:1, c9 18:1, and c9 t11 18:2) in various bovine tissues. Tissue samples were obtained from lactating Holstein cows (n=28) at slaughter, frozen in liquid nitrogen and stored at -80 °C. Total RNA was extracted and converted to complementary DNA for quantitative real time polymerase chain reaction (PCR) analysis of the SCD1 gene. Extracted lipid was converted to fatty acid methyl esters and analysed by GC. Tissues varied in expression of SCD1 gene with mammary, cardiac, intestinal adipose, and skeletal muscle expressing greater copy number as compared with lung, large intestine, small intestine and liver (371, 369, 328, 286, 257, 145, 73, and 21 copies/ng RNA, respectively). Tissues with high mRNA expression of SCD1 contained greater SCD1 protein whereas detection of SCD1 protein in tissues with low SCD1 mRNA expression was very faint or absent. Across tissues, the desaturase indices for c9 18:1 (r=0.24) and sum of SCD products (r=0.20) were positively correlated with SCD1 gene expression (P<0.01 for both). Within each tissue, the relationship between SCD1 gene expression and the desaturase indices varied. No correlation was detected between SCD1 expression and desaturase indices in the liver, large and small intestines, lung, cardiac or skeletal muscles. Positive correlations, however, were detected between SCD1 expression and the desaturase indices in intestinal adipose tissue (P<0.02 for all) except 14:1, whereas only c9 18:1, c9

  15. Corporoplasty with small soft axial prostheses (VIRILIS I®) and bovine pericardial graft (HYDRIX®) in Peyronie's disease

    PubMed Central

    Zucchi, Alessandro; Silvani, Mauro; Pecoraro, Stefano

    2013-01-01

    The surgical techniques used by Austoni and Egydio in the treatment of Peyronie's disease are based on geometric principles. The aim of this paper is to report our multicentric experience and technical changes to Austoni's original technique, focusing on several tips and tricks to make this technique easy to perform, even by less experienced practitioners. We performed operations in three different Italian institutions. We implanted a small soft Virilis I® axial prostheses (Ø 7 Fr.), using a bovine pericardium collagen matrix patch (Hydrix®) to cover the defect in the tunica albuginea. Sixty patients with a mean age of 58 years (range 44–76 years) underwent surgery between September 2005 and January 2010. After surgery, mean lengthening of the shaft was 2 cm (range 1.2–2.3 cm) with complete correction of penile recurvatum. Thirty-nine patients resumed sexual activity 60 days later, 14 after 90 days and 7 after 120 days. The international index of erectile function (IIEF) score was 15.5 before surgery and it improved to 23 at 12 and 24 months after surgery. Furthermore, the visual analogue scale (VAS) showed good results in terms of the recovery of natural sexual intercourse (over 80% of couples) and of the original length and girth of the penis. The soft implant we used takes advantage of erection that occurs spontaneously, using the residual erection of the spared cavernous tissue. The method is easy to learn and reproducible, and the use of pericardium speeds up the operation, while also covering large defects of the tunica albuginea that result from complex recurvatum. PMID:23353721

  16. Co-purification of arrestin like proteins with alpha-enolase from bovine myocardial tissues and the possible role in heart diseases as an autoantigen

    SciTech Connect

    Mirshahi, M. Le Marchand, S.

    2015-05-08

    Aim: Previously, we reported that visual arrestin co-purified with glycolytic enzymes. The aim of this study was to analyze the co-purification of arrestin like proteins (ALP) in bovine cardiac tissues with enolases. Methods: The soluble extract of bovine myocardial tissues from different regions such as left and right atriums and ventricles of the bovine heart (n = 3) was analyzed by ACA-34 gel filtration, immuno-affinity column, SDS-PAGE, ELISA, western blot and a sandwich immune assay for quantification of ALP and sequence analysis. Results: We observed that; 1) The cardiac muscle contained a 50 kDa ALP at a concentration of 751 pg/mg of soluble protein extract, 2) ALP purified, by immunoaffinity, contained alpha-enolase of 48 kDa confirmed by protein sequence analysis; 3) Cardiomyocyte cells exposed to anti arrestin and anti enolase monoclonal antibodies showed decreased proliferation in vitro, 4) High level of autoantibodies were detected by ELISA (3.57% for arrestin and 9.12% for α-enolase) in serum of patients with infarcted heart disease. Conclusion: We suggest a possible interaction between ALP and alpha-enolases yielding a complex that may be involved in the induction of cardiac autoimmune diseases. - Highlights: • We examine a possible interaction between arrestin like protein and alpha-enolases in cardiomyocyte. • We demonstrated the effect of antibodies against arrestin and enolase on cardiomyocyte cell proliferation. • We suggest that this proteins complex may be involved in the induction of cardiac autoimmune diseases.

  17. Study of laser-induced thermoelastic deformation of native and coagulated ex-vivo bovine liver tissues for estimating their optical and thermomechanical properties.

    PubMed

    Soroushian, Behrouz; Whelan, William M; Kolios, Michael C

    2010-01-01

    Several studies have explored the potential of optoacoustic imaging for monitoring thermal therapies, yet the origin of the contrast in the images is not well understood. A technique is required to measure the changes in the optical and thermomechanical properties of tissues upon coagulation to better understand this contrast. An interferometric method is presented for measuring simultaneously the optical and thermomechanical properties of native and coagulated ex-vivo bovine tissue samples based on analysis of the surface displacement of irradiated samples. Surface displacements are measured after irradiation by short laser pulses at 750 nm. A 51% decrease in the optical attenuation depth is observed for coagulated liver samples compared to native samples. No significant differences in the Grüneisen coefficient are measured in the native and coagulated tissue samples. A mean value of 0.12 for the Grüneisen coefficient is measured for both native and coagulated liver tissues. The displacement profiles exhibit consistent differences between the two tissue types. To assess the changes in the sample mechanical properties, the experimental data also are compared to numerical solutions of the equation for thermoelastic deformation. The results demonstrate that differences in the tissue expansion dynamics arise from higher values of elastic modulus for coagulated liver samples compared to native ones.

  18. Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells.

    PubMed

    Miranda, Moysés S; Nascimento, Hamilton S; Costa, Mayra P R; Costa, Nathália N; Brito, Karynne N L; Lopes, Cinthia T A; Santos, Simone S D; Cordeiro, Marcela S; Ohashi, Otávio M

    2016-10-01

    Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development. In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 10(3) and 10(4) cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 10(4) b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran). In experiment 1, co-culture with 10(4) b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 10(3) b-ATMSCs/mL (p = 0.051), group 10(4) b-ATMSCs/mL yielded higher results of blastocyst production. In experiment 2, when compared to group Gran, co-culture with 10(4) b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05). Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in

  19. Coexistence of two forms of disease-associated prion protein in extracerebral tissues of cattle infected with H-type bovine spongiform encephalopathy.

    PubMed

    Okada, Hiroyuki; Miyazawa, Kohtaro; Masujin, Kentaro; Yokoyama, Takashi

    2016-08-01

    H-type bovine spongiform encephalopathy (H-BSE) is an atypical form of BSE in aged cattle. H-BSE is characterized by the presence of two proteinase K-resistant forms of disease-associated prion protein (PrP(Sc)), identified as PrP(Sc) #1 and PrP(Sc) #2, in the brain. To investigate the coexistence of different PrP(Sc) forms in the extracerebral tissues of cattle experimentally infected with H-BSE, immunohistochemical and molecular analyses were performed by using N-terminal-, core-region- and C-terminal-specific anti-prion protein antibodies. Our results demonstrated that two distinct forms of PrP(Sc) coexisted in the various extracerebral tissues.

  20. Action of Group A Streptococcus Extracellular Product(s) on the Connective Tissue of the Bovine Heart Valve

    PubMed Central

    Goldstein, Israël; Caravano, René; Parlebas, Janine

    1974-01-01

    Group A streptococcal strains isolated from rheumatic fever patients were cultivated in the presence of bovine heart valves in a medium devoid of components from animal origin. Other group A streptococci and various bacteria were used as controls. The supernatant of these cultures was extracted and analyzed chemically and immunologically. The extracts prepared from cultures of two “rheumatogenic” strains in the presence of bovine heart valves showed fraction(s) containing proteins, oses (neutral and amined), and uronic and sialic acids. This fraction(s) was immunologically active with both anti-group A streptococcus and antisoluble connective glycoprotein antisera, with a partial identity reaction. Experiments with a diffusion chamber and attempts to precipitate a postulated enzyme from the culture of these strains by ammonium sulfate suggest that this action is due to an extracellular product of the bacteria. The meaning of these data in the physiopathology of rheumatic cardiac lesions is briefly discussed. PMID:4202889

  1. Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering

    PubMed Central

    2014-01-01

    In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly-l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly-l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation. PMID:24708858

  2. Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering

    NASA Astrophysics Data System (ADS)

    Kasálková, Nikola Slepičková; Slepička, Petr; Kolská, Zdeňka; Hodačová, Petra; Kučková, Štěpánka; Švorčík, Václav

    2014-04-01

    In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly- l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly- l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation.

  3. Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering.

    PubMed

    Kasálková, Nikola Slepičková; Slepička, Petr; Kolská, Zdeňka; Hodačová, Petra; Kučková, Stěpánka; Svorčík, Václav

    2014-04-04

    In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly-l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly-l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation.

  4. cDNA cloning of S100 calcium-binding proteins from bovine periodontal ligament and their expression in oral tissues.

    PubMed

    Duarte, W R; Kasugai, S; Iimura, T; Oida, S; Takenaga, K; Ohya, K; Ishikawa, I

    1998-09-01

    The periodontal ligament (PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. To characterize PDL cells at the molecular level, we constructed a cDNA library from bovine PDL tissue. We then focused on the isolation of S100 calcium-binding proteins (CaBPs), because they mediate Ca2+ signaling and control important cellular processes such as differentiation and metabolism. We screened the PDL cDNA library with a mouse S100A4 cDNA, and cloned the bovine cDNAs of two S100 CaBPs (S100A4 and S100A2). In northern blotting analysis, the highest expression of S100A4 was detected in PDL from erupted teeth (PDLE). PDL from teeth under eruption (PDLU) showed a lower expression of S100A4, and its expression in gingiva was faintly detectable. S100A4 expression was also high in the pulp tissue followed by the dental papilla of the tooth germ. S100A2 expression was high in PDLE and gingiva. Interestingly, only PDLE exhibited a high expression of both S100A4 and S100A2. PDLE also expressed the highest level of beta-actin, a target cytoskeletal protein for S100A4. It is conceivable that the high expression of S100A4 in PDLE is a result of the maturation of the PDL and/or a response to mechanical stress generated by mastication. Since there was a marked difference of S100A4 expression between PDL and gingiva, we propose that S100A4 could be a useful marker for distinguishing cells from these two tissues.

  5. The chemical protecting group concept applied in crosslinking of natural tissues with glutaraldehyde acetals.

    PubMed

    Goissis, G; Yoshioka, S A; Braile, D M; Ramirez, V D

    1998-03-01

    This work describes the results of the controlled crosslinking of collagen matrices by glutaraldehyde based on a double protection strategy, glutaraldehyde acetals and lysine protonation due to the acidic conditions of acetal formation. Materials crosslinked by this approach were characterized by thermal stability comparable to those obtained by conventional procedures with mechanical properties expected for bioprosthesis manufacture and with a higher stability toward collagenase hydrolysis. The integrity of the microfibrillar structure was confirmed by optical and scanning electronic microscopy. The results indicate that the glutaraldehyde acetals procedure may be of potential use for the crosslinking of bovine pericardium used in the manufacture of bioprosthetic devices. Advantages may be related to the production of materials with homogeneous crosslinking distributions, associated with better definition in the nature of the chemical link that they introduce, due to a better distribution of glutaraldehyde within the tissue matrix before the crosslinking reaction is allowed to occur. As a result, materials with improved biological and mechanical properties are expected.

  6. Propagation of tears in pericardium from young bulls: influence of the suture.

    PubMed

    Páez, José María García; Jorge-Herrero, Eduardo; Claramunt, Rafael; Millán, Isabel; Rocha, Aurora; Martínez, Belén; Cordón, Angeles; Ros, Antonio

    2010-03-01

    The tearing of the collagen fibers of biological materials utilized in implants or bioprostheses is an important, and sometimes early cause of the failure of these devices. We studied the force necessary to propagate a tear in a biomaterial, pericardium from young bulls, and the influence of the suture. An Elmendorf pendulum capable of measuring the force necessary to tear a given length of tissue was employed. We analyzed 112 trials (70%) that proved valid after achieving the homogeneity of the samples according to their thickness, thus making the results comparable. Mean forces ranging between 19.87 and 150 N were required to propagate tears measuring from 0.25 to 2.0 cm. In the samples with a 1-cm-long suture, sewn using an edge-to-edge technique, the propagation of the tear required a mean force of 15.75 N when the suture was made of nylon and 28.73 N when Prolene was utilized. When these results were compared with the mean recorded in an unsutured control series (56.76 N), the loss of resistance was significant in both sutured series (P = 0.000 and P = 0.011, respectively). Finally, the equation that relates the force (y) with the length of the tear made in unsutured tissue (x) was also obtained: y = 58.14 + 9.62x(2) (R(2) = 0.924). The force necessary to produce a microtear, thus estimated, can be utilized as a parameter for comparison.

  7. Foetal bovine intermuscular adipose tissue exhibits histological and metabolic features of brown and white adipocytes during the last third of pregnancy.

    PubMed

    Taga, H; Chilliard, Y; Picard, B; Zingaretti, M C; Bonnet, M

    2012-04-01

    This study reports the metabolic and morphological characteristics of bovine intermuscular adipose tissue (AT) throughout foetal growth. Our hypothesis was that the histological and molecular features of intermuscular AT would be different from those previously reported for foetal perirenal AT, based on its anatomical location near the muscle and the recent identification of two distinct adipocyte precursors in mouse AT depending on their locations. To address this question, intermuscular AT was sampled from Charolais and Blond d'Aquitaine foetuses at 180, 210 and 260 days post conception (dpc). The two bovine breeds were chosen because of the higher adiposity of Charolais than Blond d'Aquitaine cattle during the postnatal life. Regardless of the breed, adipocyte volume increased slightly (+38%, P < 0.01) with increasing foetal age. This was concomitant with a decrease (P < 0.05) in the activity of enzymes involved in de novo fatty acid (FA) synthesis (FA synthase and glucose-6-phosphate dehydrogenase) and FA esterification (glycerol-3-phosphate dehydrogenase) when expressed per million adipocytes, and with an increase (P ⩽ 0.01) in mRNA abundances for uncoupling protein 1, adiponectin and leptin (LEP) between 180 and 260 dpc. No difference was observed in the adipocyte volume between breeds, which was consistent with the lack of major between-breed differences in mRNA abundances or activities of enzymes involved in lipid metabolism. The mRNA abundance of lipoprotein lipase was maintained across ages, suggesting a storage of circulating FA rather than of FA synthesized de novo. Plasma LEP increased with foetal age, but only in the Charolais breed (+71%, P ⩽ 0.01), and was two- to threefold higher in Charolais than Blond d'Aquitaine foetuses. Regardless of the breed, bovine intermuscular AT contained predominantly unilocular adipocytes believed to be white adipocytes that were larger at 260 dpc than at 180 dpc. These data thus challenge current concepts of the

  8. Aortic valve reconstruction with autologous pericardium for dialysis patients.

    PubMed

    Kawase, Isamu; Ozaki, Shigeyuki; Yamashita, Hiromasa; Uchida, Shin; Nozawa, Yukinari; Matsuyama, Takayoshi; Takatoh, Mikio; Hagiwara, So

    2013-06-01

    This study aimed to report on original aortic valve reconstruction for patients on dialysis. Aortic valve reconstruction has been performed on 404 cases from April 2007 through September 2011. Among them, 54 cases on haemodialysis were retrospectively studied. Forty-seven patients had aortic stenosis, 5 had aortic regurgitation (AR), and 2 had infective endocarditis. Mean age was 70.2 ± 8.5 years. There were 35 males and 19 females. There were 27 primary aortic valve reconstructions, 11 with CABG, 6 with ascending aortic replacement, 5 with mitral valve repair and 4 with maze. First, in the procedure, harvested pericardium was treated with 0.6% glutaraldehyde solution. After resecting the cusps, we measured the distance between commissures with original sizing instrument. Then, the pericardium was trimmed with the original template. Three cusps were sutured to each annulus. Peak pressure gradient averaged to 66.0 ± 28.2 mmHg preoperatively, and decreased to 23.4 ± 10.7, 13.8 ± 5.5 and 13.3 ± 2.3 mmHg, 1 week, 1 year, and 3 years after the operation, respectively. No calcification was detected with echocardiographic follow-up. Recurrence of AR was not recorded with the mean follow-up of 847 days except for 1 case reoperated on for infective endocarditis 2.5 years after the operation. Three hospital deaths were recorded due to non-cardiac causes. Other patients were in good condition. There was no thromboembolic event. Medium-term results are excellent. Since warfarin for dialysis patients becomes problematic, a postoperative warfarin-free status is desirable. Aortic valve reconstruction can provide patients with a better quality of life without warfarin.

  9. Aortic valve reconstruction with autologous pericardium for dialysis patients†

    PubMed Central

    Kawase, Isamu; Ozaki, Shigeyuki; Yamashita, Hiromasa; Uchida, Shin; Nozawa, Yukinari; Matsuyama, Takayoshi; Takatoh, Mikio; Hagiwara, So

    2013-01-01

    OBJECTIVES This study aimed to report on original aortic valve reconstruction for patients on dialysis. METHODS Aortic valve reconstruction has been performed on 404 cases from April 2007 through September 2011. Among them, 54 cases on haemodialysis were retrospectively studied. Forty-seven patients had aortic stenosis, 5 had aortic regurgitation (AR), and 2 had infective endocarditis. Mean age was 70.2 ± 8.5 years. There were 35 males and 19 females. There were 27 primary aortic valve reconstructions, 11 with CABG, 6 with ascending aortic replacement, 5 with mitral valve repair and 4 with maze. First, in the procedure, harvested pericardium was treated with 0.6% glutaraldehyde solution. After resecting the cusps, we measured the distance between commissures with original sizing instrument. Then, the pericardium was trimmed with the original template. Three cusps were sutured to each annulus. RESULTS Peak pressure gradient averaged to 66.0 ± 28.2 mmHg preoperatively, and decreased to 23.4 ± 10.7, 13.8 ± 5.5 and 13.3 ± 2.3 mmHg, 1 week, 1 year, and 3 years after the operation, respectively. No calcification was detected with echocardiographic follow-up. Recurrence of AR was not recorded with the mean follow-up of 847 days except for 1 case reoperated on for infective endocarditis 2.5 years after the operation. Three hospital deaths were recorded due to non-cardiac causes. Other patients were in good condition. There was no thromboembolic event. CONCLUSIONS Medium-term results are excellent. Since warfarin for dialysis patients becomes problematic, a postoperative warfarin-free status is desirable. Aortic valve reconstruction can provide patients with a better quality of life without warfarin. PMID:23427312

  10. Two genes encoding the bovine mitochondrial ATP synthase proteolipid specify precursors with different import sequences and are expressed in a tissue-specific manner.

    PubMed Central

    Gay, N J; Walker, J E

    1985-01-01

    Two cDNAs encoding different precursor proteins of the same mature proteolipid subunit of mitochondrial ATP synthase have been cloned from a bovine cDNA library. The hybridisation probe was a mixture of 17-mer oligonucleotides containing 256 discrete sequences. The coding sequences of the two cDNAs differ in 25 silent positions of codons and the 3' non-coding sequences are only weakly related. The precursor sequences, which direct the import of the proteolipid into the mitochondrion, are 61 and 68 amino acids long. They are related to each other in regions which probably are recognition signals for the processing protease. The corresponding genes are expressed differently in various tissues in a way that reflects their embryonic origin. Images Fig. 3. Fig. 6. Fig. 7. PMID:2868890

  11. Increased susceptibility of transgenic mice expressing human PrP to experimental sheep bovine spongiform encephalopathy is not due to increased agent titre in sheep brain tissue

    PubMed Central

    Plinston, Chris; Hart, Patricia; Hunter, Nora; Manson, Jean C.

    2014-01-01

    Bovine spongiform encephalopathy (BSE) in cattle and variant Creutzfeldt–Jakob disease in humans have previously been shown to be caused by the same strain of transmissible spongiform encephalopathy agent. It is hypothesized that the agent spread to humans following consumption of food products prepared from infected cattle. Despite evidence supporting zoonotic transmission, mouse models expressing human prion protein (HuTg) have consistently shown poor transmission rates when inoculated with cattle BSE. Higher rates of transmission have however been observed when these mice are exposed to BSE that has been experimentally transmitted through sheep or goats, indicating that humans may potentially be more susceptible to BSE from small ruminants. Here we demonstrate that increased transmissibility of small ruminant BSE to HuTg mice was not due to replication of higher levels of infectivity in sheep brain tissue, and is instead due to other specific changes in the infectious agent. PMID:24828334

  12. Increased susceptibility of transgenic mice expressing human PrP to experimental sheep bovine spongiform encephalopathy is not due to increased agent titre in sheep brain tissue.

    PubMed

    Plinston, Chris; Hart, Patricia; Hunter, Nora; Manson, Jean C; Barron, Rona M

    2014-08-01

    Bovine spongiform encephalopathy (BSE) in cattle and variant Creutzfeldt-Jakob disease in humans have previously been shown to be caused by the same strain of transmissible spongiform encephalopathy agent. It is hypothesized that the agent spread to humans following consumption of food products prepared from infected cattle. Despite evidence supporting zoonotic transmission, mouse models expressing human prion protein (HuTg) have consistently shown poor transmission rates when inoculated with cattle BSE. Higher rates of transmission have however been observed when these mice are exposed to BSE that has been experimentally transmitted through sheep or goats, indicating that humans may potentially be more susceptible to BSE from small ruminants. Here we demonstrate that increased transmissibility of small ruminant BSE to HuTg mice was not due to replication of higher levels of infectivity in sheep brain tissue, and is instead due to other specific changes in the infectious agent. © 2014 The Authors.

  13. Comparison of whole body and tissue blood volumes in rainbow trout (Salmo gairdneri) with 125I bovine serum albumin and 51Cr-erythrocyte tracers

    USGS Publications Warehouse

    Gingerich, W.H.; Pityer, R.A.

    1989-01-01

    Total, packed cell and, plasma volume estimates were made for the whole body and selected tissues of rainbow trout by the simultaneous injection of radiolabelled trout erythrocyte (51Cr-RBC) and radioiodinated bovine serum albumin (125I-BSA) tracers. Blood volumes were estimated with both markers separately by the tracer-hematocrit method and as the combination of the 51Cr-RBC packed cell and 125I-BSA plasma volumes. Mean whole body blood volume was significantly less when calculated from the 51Cr-RBC tracer data (3.52±0.78 ml/100 g; ±SD) than when calculated with the 125I-BSA tracer (5.06±0.86 ml/100 g) or as the sum of the two volumes combined (4.49±0.60 ml/100 g). The whole body hematocrit (28±5%), estimated as the quotient of the 51Cr-RBC volume divided by the sum of the 125I-BSA and the 51Cr-RBC volumes, also was significantly less than the dorsal aortic microhematocrit (36±4%). Estimates of total blood volumes in most tissues were significantly smaller when calculated from the51Cr-RBC data than when calculated by the other two methods. Tissue blood volumes were greatest in highly vascularized and well perfused tissues and least in poorly vascularized tissues. The relative degree of vascularization among tissues generally remained the same regardless of whether the red cell or the plasma tracer was used to calculated blood volume. It is not clear whether the expanded plasma volume is the result of the distribution of erythrocyte-poor blood into the secondary circulation or the result of extravascular exchange of plasma proteins.

  14. Effects of cell culture techniques on gene expression and cholesterol efflux in primary bovine mammary epithelial cells derived from milk and tissue.

    PubMed

    Sorg, D; Potzel, A; Beck, M; Meyer, H H D; Viturro, E; Kliem, H

    2012-10-01

    Primary bovine mammary epithelial cells (pbMEC) are often used in cell culture to study metabolic and inflammatory processes in the udder of dairy cows. The most common source is udder tissue from biopsy or after slaughter. However, it is also possible to culture them from milk, which is non-invasive, repeatable and yields less contamination with fibroblasts. Generally, not much is known about the influence of cell origin and cell culture techniques such as cryopreservation on pbMEC functionality. Cells were extracted from milk and udder tissue to evaluate if milk-derived pbMEC are a suitable alternative to tissue-derived pbMEC and to test what influence cryopreservation has. The cells were cultivated for three passages and stored in liquid nitrogen. The relative gene expression of the five target genes kappa-casein, lingual antimicrobial peptide (LAP), lactoferrin, lysozyme (LYZ1) and the prolactin receptor normalised with keratin 8 showed a tendency to decrease in the tissue cultures, but not in the milk-derived cultures, suggesting a greater influence of the cultivation process on tissue-derived cells, freezing lowered expression levels in both cultures. Overall expression of LAP and LYZ1 tended to be higher in milk cells. Cholesterol efflux was measured to compare passages one to seven in milk-derived cells. Passage number did not alter the efflux rate (p ≤ 0.05). We showed for the first time that the extraction of pbMEC from milk can be a suitable alternative to tissue extraction.

  15. Bovine somatotropin and rumen-undegradable protein effects in prepubertal dairy heifers: effects on body composition and organ and tissue weights.

    PubMed

    Moallem, U; Dahl, G E; Duffey, E K; Capuco, A V; Wood, D L; McLeod, K R; Baldwin, R L; Erdman, R A

    2004-11-01

    The objectives of this study were to determine the effect of recombinant bovine somatotropin (bST) and added dietary rumen undegradable protein (RUP) on organ and tissue weights and body composition in growing dairy heifers. Thirty-two Holstein heifers were in the experiment, 8 killed initially at 3 mo of age, with the remaining 24 Holstein heifers randomly assigned to treatments (n = 6) consisting of 0.1 mg/kg of body weight per day of bST and 2% added dietary RUP (dry matter basis) applied in a 2 x2 factorial design. A total of 6 heifers per treatment group (3 each at 5 and 10 mo of age), were slaughtered to determine body composition and organ masses. Feed intake measured from group intakes were increased by 0.25 and 0.35 kg/d with bST and RUP, respectively. Administration of bST tended to increase the weights of visceral organs including heart, kidney, and spleen by 16, 16, and 38%, respectively. At 10 mo of age, there was a trend for increased empty body weights (EBW) and non-carcass components for heifers treated with bST, but there were no effects of RUP. Body components and organ weights, expressed as a percentage of BW were not affected by RUP or bST. Somatotropin increased ash weight at 10 mo without affecting amounts of protein, fat, and energy. Rates of ash deposition between 3 and 10 mo of age were increased 7 and 4 g/d by bST and RUP, respectively. There were no treatment effects on rates of body fat, protein, and energy deposition. Bovine somatotropin and RUP altered the metabolism of growing heifers in a manner that was consistent with increased rates of skeletal growth. This suggests that nutritional and endocrine manipulations could increase growth rates of skeletal tissues without increasing fat deposition in prepubertal dairy heifers.

  16. Comparison between immunohistochemistry and two PCR methods for detection of Neospora caninum in formalin-fixed and paraffin-embedded brain tissue of bovine fetuses.

    PubMed

    Sánchez, G F D; Banda, R V M; Sahagun, R A; Ledesma, M N; Morales, S E

    2009-10-14

    The objective of this study was to identify the presence of the parasite by comparing immunohistochemistry (IHC) with two polymerase chain reaction (PCR) methods for the detection of the pNc5 gene and the internal transcribed spacer 1 (ITS1) of N. caninum in brain tissue of bovine fetuses that had previously been fixed in formalin and paraffin-embedded. In 29 out of 48 brains (60.4%), microscopic lesions consistent with Neospora infection were observed, and 21 of the 29 cases (72.41%) were positive for IHC. Fifteen of the 29 cases positive for IHC (51.72%) were also positive on the ITS1 PCR, and 12 cases were also positive on the pNc5 PCR (41.37%). The sensitivity of the PCR assays was 71.42% and 57.14%, respectively, and the specificity was 100% for both. The concordance between histopathology and IHC and the ITS1 PCR was 85%, and in the case of the pNc5 PCR it was 77.5%. When the number of fetuses positive by IHC and both PCR tests was compared, no statistically significant difference was found (P>0.05). It is concluded that the use of formalin-fixed and paraffin-embedded bovine fetal tissues allows the detection of N. caninum by IHC or PCR. Nevertheless, it is recommended that more than one technique is used to increase the diagnostic sensitivity, and preferably tests that show better performance in the individual laboratory should be selected.

  17. Effect of the anatomical site on telomere length and pref-1 gene expression in bovine adipose tissues

    SciTech Connect

    Yamada, Tomoya Higuchi, Mikito; Nakanishi, Naoto

    2015-08-07

    Adipose tissue growth is associated with preadipocyte proliferation and differentiation. Telomere length is a biological marker for cell proliferation. Preadipocyte factor-1 (pref-1) is specifically expressed in preadipocytes and acts as a molecular gatekeeper of adipogenesis. In the present study, we investigated the fat depot-specific differences in telomere length and pref-1 gene expression in various anatomical sites (subcutaneous, intramuscular and visceral) of fattening Wagyu cattle. Visceral adipose tissue expressed higher pref-1 mRNA than did subcutaneous and intramuscular adipose tissues. The telomere length in visceral adipose tissue tended to be longer than that of subcutaneous and intramuscular adipose tissues. The telomere length of adipose tissue was not associated with adipocyte size from three anatomical sites. No significant correlation was found between the pref-1 mRNA level and the subcutaneous adipocyte size. In contrast, the pref-1 mRNA level was negatively correlated with the intramuscular and visceral adipocyte size. These results suggest that anatomical sites of adipose tissue affect the telomere length and expression pattern of the pref-1 gene in a fat depot-specific manner. - Highlights: • Visceral adipose tissue express higher pref-1 mRNA than other anatomical sites. • Telomere length in visceral adipose tissue is longer than other anatomical sites. • Telomere length of adipose tissue is not associated with adipocyte size. • Pref-1 mRNA is negatively correlated with intramuscular and visceral adipocyte size.

  18. Micro-mechanical model for the tension-stabilized enzymatic degradation of collagen tissues

    NASA Astrophysics Data System (ADS)

    Nguyen, Thao; Ruberti, Jeffery

    We present a study of how the collagen fiber structure influences the enzymatic degradation of collagen tissues. Experiments of collagen fibrils and tissues show that mechanical tension can slow and halt enzymatic degradation. Tissue-level experiments also show that degradation rate is minimum at a stretch level coincident with the onset of strain-stiffening in the stress response. To understand these phenomena, we developed a micro-mechanical model of a fibrous collagen tissue undergoing enzymatic degradation. Collagen fibers are described as sinusoidal elastica beams, and the tissue is described as a distribution of fibers. We assumed that the degradation reaction is inhibited by the axial strain energy of the crimped collagen fibers. The degradation rate law was calibrated to experiments on isolated single fibrils from bovine sclera. The fiber crimp and properties were fit to uniaxial tension tests of tissue strips. The fibril-level kinetic and tissue-level structural parameters were used to predict tissue-level degradation-induced creep rate under a constant applied force. We showed that we could accurately predict the degradation-induce creep rate of the pericardium and cornea once we accounted for differences in the fiber crimp structure and properties.

  19. Imaging the pericardium: appearances on ECG-gated 64-detector row cardiac computed tomography

    PubMed Central

    O'Leary, S M; Williams, P L; Williams, M P; Edwards, A J; Roobottom, C A; Morgan-Hughes, G J; Manghat, N E

    2010-01-01

    Multidetector row computed tomography (MDCT) with its high spatial and temporal resolution has now become an established and complementary method for cardiac imaging. It can now be used reliably to exclude significant coronary artery disease and delineate complex coronary artery anomalies, and has become a valuable problem-solving tool. Our experience with MDCT imaging suggests that it is clinically useful for imaging the pericardium. It is important to be aware of the normal anatomy of the pericardium and not mistake normal variations for pathology. The pericardial recesses are visible in up to 44% of non-electrocardiogram (ECG)-gated MDCT images. Abnormalities of the pericardium can now be identified with increasing certainty on 64-detector row CT; they may be the key to diagnosis and therefore must not be overlooked. This educational review of the pericardium will cover different imaging techniques, with a significant emphasis on MDCT. We have a large research and clinical experience of ECG-gated cardiac CT and will demonstrate examples of pericardial recesses, their variations and a wide variety of pericardial abnormalities and systemic conditions affecting the pericardium. We give a brief relevant background of the conditions and reinforce the key imaging features. We aim to provide a pictorial demonstration of the wide variety of abnormalities of the pericardium and the pitfalls in the diagnosis of pericardial disease. PMID:20197434

  20. On the Applicability of the Thermal Dose Cumulative Equivalent Minutes Metric to the Denaturation of Bovine Serum Albumin in a Polyacrylamide Tissue Phantom

    NASA Astrophysics Data System (ADS)

    Nandlall, Sacha D.; Arora, Manish; Schiffter, Heiko A.; Coussios, Constantin-C.

    2009-04-01

    Thermal dose has been proposed for various hyperthermic cancer treatment modalities as a measure of heat-induced tissue damage. However, the applicability of current thermal dose metrics to tissue is not well understood, particularly at the temperatures and rates of heating relevant to ablative cancer therapy using High-Intensity Focussed Ultrasound (HIFU). In this work, we assess whether the most widely employed thermal dose metric, Cumulative Equivalent Minutes (CEM), can adequately quantify heat-induced denaturation in a tissue-mimicking material (phantom) consisting of Bovine Serum Albumin (BSA) proteins embedded in a polyacrylamide matrix. The phantom is exposed to various temperature profiles and imaged under controlled lighting conditions against a black background as it denatures and becomes progressively more opaque. Under the assumption that the mean backscattered luminous intensity provides a good measure of the extent of BSA denaturation, we establish a relationship between the amount of thermal damage caused to the phantom, exposure time, and temperature. We demonstrate that, for monotonically increasing and bounded temperature profiles, the maximal degree to which the phantom can denature is dependent on the peak temperature it reaches, irrespective of exposure duration. We also show that when the CEM is computed using the commonly employed piecewise-constant approximation of the parameter R, the CEM values corresponding to the same degree of damage delivered using different temperature profiles do not agree well with each other in general.

  1. On the Applicability of the Thermal Dose Cumulative Equivalent Minutes Metric to the Denaturation of Bovine Serum Albumin in a Polyacrylamide Tissue Phantom

    SciTech Connect

    Nandlall, Sacha D.; Arora, Manish; Schiffter, Heiko A.; Coussios, Constantin-C.

    2009-04-14

    Thermal dose has been proposed for various hyperthermic cancer treatment modalities as a measure of heat-induced tissue damage. However, the applicability of current thermal dose metrics to tissue is not well understood, particularly at the temperatures and rates of heating relevant to ablative cancer therapy using High-Intensity Focussed Ultrasound (HIFU). In this work, we assess whether the most widely employed thermal dose metric, Cumulative Equivalent Minutes (CEM), can adequately quantify heat-induced denaturation in a tissue-mimicking material (phantom) consisting of Bovine Serum Albumin (BSA) proteins embedded in a polyacrylamide matrix. The phantom is exposed to various temperature profiles and imaged under controlled lighting conditions against a black background as it denatures and becomes progressively more opaque. Under the assumption that the mean backscattered luminous intensity provides a good measure of the extent of BSA denaturation, we establish a relationship between the amount of thermal damage caused to the phantom, exposure time, and temperature. We demonstrate that, for monotonically increasing and bounded temperature profiles, the maximal degree to which the phantom can denature is dependent on the peak temperature it reaches, irrespective of exposure duration. We also show that when the CEM is computed using the commonly employed piecewise-constant approximation of the parameter R, the CEM values corresponding to the same degree of damage delivered using different temperature profiles do not agree well with each other in general.

  2. Aortic valve replacement with the Cardioprotese Premium bovine pericardium bioprosthesis: four-year clinical results

    PubMed Central

    Farias, Fábio Rocha; da Costa, Francisco Diniz Affonso; Balbi Filho, Eduardo Mendel; Fornazari, Daniele de Fátima; Collatusso, Claudinei; Ferreira, Andreia Dumsch de Aragon; Lopes, Sergio Veiga; Fernandes, Tadeu Augusto

    2012-01-01

    OBJECTIVES This study reports the initial clinical and echocardiographic results of the Premium bioprosthetic aortic valve up to 4 years of follow-up. METHODS Between October 2007 and July 2011, 121 consecutive patients were submitted for aortic valve replacement with the Premium bioprosthetic valve. The mean age was 68 ± 9 years and 64 patients were males. The patients were periodically evaluated by clinical and echocardiographic examinations. The mean follow-up was 21 months (min = 2, max = 48), yielding 217 patients/year for the analysis. RESULTS The hospital mortality was 8%. Late survival at 3 years was 89% (95% CI: 81.9–93.3%), and 80% of the patients were in NYHA functional class I/II. The rates of valve-related complications were low, with a linearized incidence of 0.9%/100 patients/year for thromboembolic complications, 0% for haemorrhagic events and 0.9%/100 patients/year of bacterial endocarditis. There was no case of primary structural valve dysfunction. The mean effective orifice area was 1.61 ± 0.45 cm2; mean gradient 13 ± 5 mmHg and peak gradient 22 ± 9 mmHg. Significant patient–prosthesis mismatch was found in only 11% of the cases. CONCLUSIONS The Premium bioprosthetic aortic valve demonstrated very satisfactory clinical and echocardiographic results up to 4 years, similar to other commercially available, third-generation bioprosthetic valves. PMID:22588029

  3. Localized ridge defect augmentation using human pericardium membrane and demineralized bone matrix.

    PubMed

    Vidyadharan, Arun Kumar; Ravindran, Anjana

    2014-01-01

    Patient wanted to restore her lost teeth with implants in the lower left first molar and second premolar region. Cone beam computerized tomography (CBCT) revealed inadequate bone width and height around future implant sites. The extraction socket of second premolar area revealed inadequate socket healing with sparse bone fill after 4 months of extraction. To evaluate the clinical feasibility of using a collagen physical resorbable barrier made of human pericardium (HP) to augment localized alveolar ridge defects for the subsequent placement of dental implants. Ridge augmentation was done in the compromised area using Puros® demineralized bone matrix (DBM) Putty with chips and an HP allograft membrane. Horizontal (width) and vertical hard tissue measurements with CBCT were recorded on the day of ridge augmentation surgery, 4 month and 7 months follow-up. Intra oral periapical taken 1 year after implant installation showed minimal crestal bone loss. Bone volume achieved through guided bone regeneration was a gain of 4.8 mm horizontally (width) and 6.8 mm vertically in the deficient ridge within a period of 7 months following the procedure. The results suggested that HP Allograft membrane may be a suitable component for augmentation of localized alveolar ridge defects in conjunction with DBM with bone chips.

  4. Effect of the anatomical site on telomere length and pref-1 gene expression in bovine adipose tissues.

    PubMed

    Yamada, Tomoya; Higuchi, Mikito; Nakanishi, Naoto

    2015-08-07

    Adipose tissue growth is associated with preadipocyte proliferation and differentiation. Telomere length is a biological marker for cell proliferation. Preadipocyte factor-1 (pref-1) is specifically expressed in preadipocytes and acts as a molecular gatekeeper of adipogenesis. In the present study, we investigated the fat depot-specific differences in telomere length and pref-1 gene expression in various anatomical sites (subcutaneous, intramuscular and visceral) of fattening Wagyu cattle. Visceral adipose tissue expressed higher pref-1 mRNA than did subcutaneous and intramuscular adipose tissues. The telomere length in visceral adipose tissue tended to be longer than that of subcutaneous and intramuscular adipose tissues. The telomere length of adipose tissue was not associated with adipocyte size from three anatomical sites. No significant correlation was found between the pref-1 mRNA level and the subcutaneous adipocyte size. In contrast, the pref-1 mRNA level was negatively correlated with the intramuscular and visceral adipocyte size. These results suggest that anatomical sites of adipose tissue affect the telomere length and expression pattern of the pref-1 gene in a fat depot-specific manner.

  5. Evaluation of a tissue-engineered bovine pericardial patch in paediatric patients with congenital cardiac anomalies: initial experience with the ADAPT-treated CardioCel® patch

    PubMed Central

    Neethling, William M.L.; Strange, Geoff; Firth, Laura; Smit, Francis E.

    2013-01-01

    OBJECTIVES This study evaluated the safety, efficacy and clinical performance of the tissue-engineered ADAPT® bovine pericardial patch (ABPP) in paediatric patients with a range of congenital cardiac anomalies. METHODS In this single-centre, prospective, non-randomized clinical study, paediatric patients underwent surgery for insertion of the ABPP. Primary efficacy measures included early (<30 day) morbidity; incidence of device-related complications; haemodynamic performance derived from echocardiography assessment at 6- and 12-month follow-up and magnetic resonance imaging findings in 10 randomly selected patients at 12 months. Secondary measures included device-handling characteristics; shape and sizing characteristics and perioperative implant complications. The Aristotle complexity scoring system was used to score the complexity level of all surgical procedures. Patients completing the 12-month study were eligible to enter a long-term evaluation study. RESULTS Between April 2008 and September 2009, the ABPP was used in 30 paediatric patients. In the 30-day postoperative period, no graft-related morbidity was observed. In total, there were 5 deaths (2 in the 30-day postoperative period and 3 within the first 6 postoperative months). All deaths were deemed due to comorbid non-graft-related events. Echocardiography assessment at 6 and 12 months revealed intact anatomical and haemodynamically stable repairs without any visible calcification of the patch. Magnetic resonance imaging assessment in 10 patients at 12 months revealed no signs of calcification. Fisher's exact test demonstrated that patients undergoing more complex, higher risk surgical repairs (Aristotle complexity score >8) were significantly more likely to die (P = 0.0055, 58% survival compared with 100% survival for less complex surgical repairs). In 19 patients, echocardiographic data were available at 18–36 months with no evidence of device calcification, infection, thromboembolic events or

  6. Tissue-specific effects of in vitro fertilization procedures on genomic cytosine methylation levels in overgrown and normal sized bovine fetuses.

    PubMed

    Hiendleder, Stefan; Wirtz, Michaela; Mund, Cora; Klempt, Martina; Reichenbach, Horst-Dieter; Stojkovic, Miodrag; Weppert, Myriam; Wenigerkind, Hendrik; Elmlinger, Martin; Lyko, Frank; Schmitz, Oliver J; Wolf, Eckhard

    2006-07-01

    Epigenetic perturbations are assumed to be responsible for phenotypic abnormalities of fetuses and offspring originating from in vitro embryo techniques. We studied 29 viable Day-80 bovine fetuses to assess the effects of two in vitro fertilization protocols (IVF1 and IVF2) on fetal phenotype and genomic cytosine methylation levels in liver, skeletal muscle, and brain. The IVF1 protocol employed 0.01 U/ml of FSH and LH in oocyte maturation medium and 5% estrous cow serum (ECS) in embryo culture medium, whereas the IVF2 protocol employed 0.2 U/ml of FSH and no LH for oocyte maturation and 10% ECS for embryo culture. Comparisons with in vivo-fertilized controls (n=14) indicated an apparently normal phenotype for IVF1 fetuses (n=5), but IVF2 fetuses (n=10) were significantly heavier (19.9%) and longer (4.7%), with increased heart (25.2%) and liver (27.9%) weights, and thus displayed an overgrowth phenotype. A clinicochemical screen of 18 plasma parameters revealed significantly increased levels of insulin-like growth factor 1 (40.8%) and creatinine (37.5%) in IVF2, but not in IVF1, fetuses. Quantification of genomic 5-methylcytosine (5mC) by capillary electrophoresis indicated that both IVF1 and IVF2 fetuses differed from controls. We observed significant DNA hypomethylation in liver and muscle of IVF1 fetuses (-16.1% and -9.3%, respectively) and significant hypermethylation in liver of IVF2 fetuses (+11.2%). The 5mC level of cerebral DNA was not affected by IVF protocol. Our data indicate that bovine IVF procedures can affect fetal genomic 5mC levels in a protocol- and tissue-specific manner and show that hepatic hypermethylation is associated with fetal overgrowth and its correlated endocrine changes.

  7. Tissue-specific elevated genomic cytosine methylation levels are associated with an overgrowth phenotype of bovine fetuses derived by in vitro techniques.

    PubMed

    Hiendleder, Stefan; Mund, Cora; Reichenbach, Horst-Dieter; Wenigerkind, Hendrik; Brem, Gottfried; Zakhartchenko, Valeri; Lyko, Frank; Wolf, Eckhard

    2004-07-01

    Epigenetic perturbations are assumed to be responsible for abnormalities observed in fetuses and offspring derived by in vitro techniques. We have designed an experiment with bovine Day 80 fetuses generated by somatic cell nuclear transfer (SCNT), in vitro fertilization (IVF), and artificial insemination (AI) to determine the relationship between fetal phenotype and genome-wide 5-methylcytosine (5mC) content. When compared with AI controls, SCNT and IVF fetuses displayed significantly increased body weight (61% and 28%), liver weight (100% and 36%), and thorax circumference (20% and 11%). A reduced crown-rump length:thorax circumference ratio (1.175 +/- 0.017 in SCNT and 1.292 +/- 0.018 in IVF vs. 1.390 +/- 0.018 in AI, P < 0.001 and P < 0.002) was the external hallmark of this disproportionate overgrowth phenotype. The SCNT fetuses showed significant hypermethylation of liver DNA in comparison with AI controls (3.46% +/- 0.08% vs. 3.17% +/- 0.09% 5mC, P < 0.03), and the cytosine methylation levels for IVF fetuses (3.34% +/- 0.09%) were, as observed for phenotypic parameters, intermediate to the other groups. Regressions of fetal body and liver weight and thorax circumference on 5mC content of liver DNA were positive (P < 0.073-0.079). Furthermore, a significant negative regression (P < 0.021) of the crown-rump length:thorax circumference ratio on liver 5mC was observed. The 5mC content of placental cotyledon DNA was 46% lower than in liver DNA (P < 0.0001) but did not differ among groups. These data are in striking contrast with the recently reported hypomethylation of DNA from SCNT fetuses and indicate that hypermethylation of fetal tissue, but not placenta, is linked to the overgrowth phenotype in bovine SCNT and IVF fetuses.

  8. Screening of danofloxacin residue in bovine tissue by terbium-sensitized luminescence on c18 sorbent strips.

    PubMed

    Chen, Guoying; Du, Yue

    2011-02-23

    Danofloxacin (DANO) residue in bovine muscle was screened at 200 ng/g by terbium-sensitized luminescence (TSL) directly measured on 10 × 6 mm C18 sorbent strips. The analyte was first adsorbed on sorbent surface by immersion in defatted homogenates. After reagent application and desiccation, TSL was directly measured on sorbent surfaces at λ(ex) = 273 nm and λ(em) = 546 nm. The luminescence intensity was linearly dependent on DANO concentration in the 0-1000 ng/g range (R(2) = 0.9967). A threshold was established at x(200) - 3σ(200), where x(200) and σ(200) are the mean and standard deviation, respectively, of the DANO signals at 200 ng/g. Among 48 blind samples randomly fortified at 0-1000 ng/g, 45 were screened correctly and 3 negative samples were presumed positive. This simple screening protocol has the potential to significantly reduce sample numbers and hence improve sample throughput and save assay costs.

  9. Characterization and phylogenetic analysis of Bovine viral diarrhea virus in brain tissues from nonambulatory (downer) cattle in Korea.

    PubMed

    Oem, Jae-Ku; Chung, Joon-Yee; Roh, In-Soon; Kim, Hye-Ryoung; Bae, You-Chan; Lee, Kyung-Hyun; Jin, Young-Hwa; Lee, O-Soo

    2010-07-01

    Between August 2008 and May 2009, 386 brain and serum samples from adult cattle (2-7 years old) showing a variety of clinical signs of downer cow syndrome were received by the National Veterinary Research and Quarantine Service. All brain samples were tested for the presence of Bovine viral diarrhea virus (BVDV) by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and antigen capture ELISA (Ag-ELISA). The BVDV nucleic acid was detected in 54 of 386 (15.5%) brain samples tested by RT-PCR. Positive results were detected in 14 (3.67%) and 13 (3.4%) of samples tested by IHC and Ag-ELISA, respectively. Both BVDV nucleic acid and antigen were detected in 11 cattle (2.9%) by all 3 diagnostic tests; however, antibodies against BVDV were not detected in these 11 cattle. A molecular classification of the identified viral strains (n = 40) was also carried out. Neighbor-joining phylogenetic analysis revealed that most of the identified viruses belonged to BVDV genotype 1a (n = 10), 1b (n = 16), and 2a (n = 8). The remaining strains were subtypes 1c (n = 1), 1n (n = 4), and 1m (n = 1). Interestingly, most of the BVDV-1b strains (n = 9) identified in brain samples were confirmed by all 3 diagnostic tests. Further studies should be performed to determine why the BVDV-1b strain was found in brain samples that were positive using all 3 diagnostic tests.

  10. Inactivation of foot-and-mouth disease virus in various bovine tissues used for the production of natural sausage casings.

    PubMed

    Wijnker, Joris J; Haas, Bernd; Berends, Boyd R

    2012-02-01

    Bovine intestines, bladders and oesophagus are used for the production of natural casings ("beef casings") as edible sausage containers. Derived from cattle experimentally infected with FMDV (initial dosage 10(4) TCID(50)/mL, strain A Iran 97), these beef casings were treated with sodium chloride (NaCl) or phosphate supplemented salt (P-salt). In addition, different in-vitro experiments using beef casings were done on a small scale with other FMDV strains (A Turkey 06, C-Oberbayern and O(1) Manisa) as "proof of principle". Based on the combined results of the in-vivo and in-vitro experiments, it can be concluded that the storage period of 30 days at 20 °C in NaCl is sufficiently effective to inactivate a possible contamination with FMDV in beef casings and that the usage of P-salt does not clearly enhance the inactivation of FMDV infectivity. Storage of salted beef casings at about 20 °C for 30 days is already part of the Standard Operating Procedures (included in HACCP) of the international casing industry and can therefore be considered as a protective measure for the international trade in natural casings.

  11. Development and validation of a highly sensitive quantitative/confirmatory method for the determination of ivermectin residues in bovine tissues by UHPLC-MS/MS.

    PubMed

    Hoyos O, Duvan E; Cuartas O, Yudy A; Peñuela M, Gustavo A

    2017-04-15

    In this work, a highly sensitive quantitative/confirmatory method was developed for the selective determination of marker residue 22,23-dihydroavermectin B1a in fat, kidney, liver and muscle bovine tissues using UHPLC-MS/MS. A QuEChERS method and a subsequent d-SPE cleaning with MgSO4 and C18 were used for samples preparation. The method was validated at three levels in each tissue (8.0, 20.0 and 80.0μg/kg for fat, kidney and muscle, 8.0, 16.0 and 40.0μg/kg for liver), taking into account international guidelines. Linearity and matrix effect, accuracy, within-laboratory repeatability, specificity, LOQ, LOD, Q/q relative ion intensities and uncertainty were reported. Recoveries were obtained in the range of 70-120% with within-laboratory repeatability less than 20% for the overall working range. LODs varied from 0.05 to 0.12μg/kg. Compound identification was performed using the relative intensities of monitored transitions. The method could be applied to monitoring withdrawal period specified for ivermectin in cattle.

  12. Topographic distribution of the different cell types, connective tissue and vascular tissue/lumina within a functional bovine corpus luteum and its association with breed, type of fixation protocol and stage during the cycle.

    PubMed

    Cools, S; Van den Broeck, W; De Vliegher, S; Piepers, S; Hostens, M; Opsomer, G

    2013-08-01

    In the present study, we analysed the effect of fixative, breed, luteal stage and location on the nuclear density, volume density of connective tissue and vascular tissue/lumina within a bovine luteal gland in view of the development of an in vivo sampling technique to longitudinally monitor luteal histophysiology. The inner zone defined as the zone geometrically closest to the centre of the gland shows a significantly lower nuclear density (for all cell types) and a higher volume density of collagen fibres and vessels when compared with the outer zone (p < 0.001). The nuclear density in luteal glands from Holstein-Friesian cows is not significantly different from that in Belgian Blue cows, nor is it in stage II vs stage III glands. The collagen fibre content was significantly lower in glands of Belgian Blue cows (p = 0.01) and in younger glands (p = 0.003). Hence, it seems that the lower nuclear density in the inner zone was compensated by a higher amount of collagen fibres. As the type of fixative applied has a significant effect on the nuclear density of the different cell types, the present study warrants future research to further optimize the fixation protocol. As a conclusion, we can state that the topographic difference in nuclear distribution for the different cell types in a bovine luteal gland is only significant when comparing the inner vs the outer zone. This implies that if a sample representative for the whole gland has to be taken, for example, when taking an in vivo sample, it is necessary that the biopsy goes through the inner zone and contains the total diameter of the gland.

  13. Comparison of effectiveness and safety of operations on the pericardium

    SciTech Connect

    Palatianos, G.M.; Thurer, R.J.; Kaiser, G.A.

    1985-07-01

    A ten-year experience with operations on the pericardium in 71 consecutive patients was reviewed. The patients ranged in age from nine months to 75 years old. Fifty-three patients were operated upon for pericardial effusion and 14 for pericardial constriction. Sixty-seven patients had pericarditis: 21 of them underwent subxiphoid tube drainage; ten, limited pericardiectomy; and the remaining 36, extensive pericardiectomy. There were two trauma victims who underwent diagnostic pericardiotomy. Two patients underwent excision of pericardial cysts. There was one operative death during extensive pericardiectomy for constrictive tuberculous pericarditis. Thirty-day mortality was three of 21 patients after subxiphoid tube drainage, three of ten after limited pericardiectomy and five of 36 after extensive pericardiectomy. Postoperative complications consisted of pulmonary problems in four patients (two after pericardiectomy and two after extensive pericardiectomy), cardiac arrhythmia in one patient after tube drainage and postpericardiotomy syndrome in one patient after extensive pericardiectomy. Mean follow-up was 3.2 years. Recurrent pericardial effusion occurred in two patients; one had limited pericardiectomy but did not require reoperation and one had diagnostic pericardiotomy without drainage. Six patients with persistent postirradiation pericardial effusion were treated effectively with extensive pericardiotomy. Experience indicates that subxiphoid tube drainage is effective for the treatment of pericardial effusion and safer than limited pericardiectomy, while extensive pericardiectomy is the operation of choice for pericardial constriction and radiation-related pericardial effusion.

  14. Detection of bovine Herpesvirus type 5 in formalin-fixed, paraffin-embedded bovine brain by PCR: a useful adjunct to conventional tissue-based diagnostic test of bovine encephalitis.

    PubMed

    Ferrari, H F; Luvizotto, M C R; Rahal, P; Cardoso, T C

    2007-12-01

    The aim of this study was to evaluate the application of PCR technique for the detection of BoHV-5 in routinely formalin-fixed, paraffin-embedded brain tissues in 20 naturally infected calves affected by fatal meningoencephalitis. Brains were divided into two halves, one kept fresh for virus isolation and PCR assay, targeting the glycoprotein C gene from BoHV-5 genome. The other half brain, corresponding to posterior cortex region, was submitted to formalin fixation and embedded into paraffin blocks for microscopic evaluation and total DNA isolation. Most of the slides showed severe multifocal non-supurative encephalitis with neuronal degeneration, neurophagia, and no acidophilic intranuclear inclusions could be found in neurons and glial. The 20 fresh samples were confirmed, by virus isolation and PCR assay, as having the BoHV-5 virus and, respective glicoprotein C sequence, while 15 of 20 formalin-fixed, paraffin-embedded samples were considered positive for the same analysis. The results revealed the first description of PCR efficiency, applied to formalin-fixed, paraffin-embedded brain collected from naturally infected calves, improving the detection of BoHV-5 from archival samples in South America.

  15. Distribution pattern of bovine viral diarrhoea virus type 1 genome in lymphoid tissues of experimentally infected sheep.

    PubMed

    Karikalan, M; Rajukumar, K; Mishra, N; Kumar, M; Kalaiyarasu, S; Rajesh, K; Gavade, V; Behera, S P; Dubey, S C

    2016-06-01

    In this study, cellular localization and the distribution pattern of BVDV genome in lymphoid tissues during the course of experimental acute BVDV-1 infection of sheep was investigated. Tonsils, mesenteric lymph nodes (MLN) and spleen were collected on 3, 6, 9, 12 and 15 days post infection (dpi) from twenty 4-month-old lambs, experimentally inoculated intra-nasally with 5 × 10(5) TCID50 of a non-cytopathic (ncp) BVDV-1 isolate, Ind-17555. Tissues collected from ten mock-infected lambs served as controls. In situ hybridization (ISH) was carried out in paraformaldehyde fixed paraffin embedded tissue sections using digoxigenin labelled riboprobe targeting 5'-UTR of BVDV-1. BVDV genome was detected at all the intervals from 3 dpi to 15 dpi in the lymphoid tissues with variations between the intervals and also amongst the infected sheep. During the early phase of acute infection, presence of viral genome was more in tonsils than MLN and spleen, whereas the distribution was higher in MLN during later stages. BVDV-1 genome positive cells included lymphocytes, macrophages, plasma cells, reticular cells and sometimes crypt epithelial cells. Genome distribution was frequently observed in the lymphoid follicles of tonsils, MLN and spleen, besides the crypt epithelium in tonsils, paracortex and medullary sinus and cords of MLN. Most abundant and widespread distribution of BVDV-1 genome was observed on 6 dpi while there was a reduction in number and intensity of positive signals by 15 dpi in most of the infected animals. This is the first attempt made to study the localisation of BVDV-1 in lymphoid tissues of acutely infected sheep by in situ hybridization. The results show that the kinetics of BVDV-1 distribution in lymphoid tissues of experimentally infected non-pregnant sheep follows almost a similar pattern to that demonstrated in BVDV infected cattle.

  16. Effect on hematopoietic tissue of experimental infection of calves with noncytopathic type 2 bovine viral diarrhea virus

    PubMed Central

    2004-01-01

    Abstract To investigate the hematologic abnormalities observed with noncytopathic type 2 bovine viral diarrhea virus (ncpBVDV-2), calves 6 to 8 mo old were inoculated with an isolate of either high virulence (HV24515) or low virulence (LV11Q); control animals received the same volume of uninfected cell-culture supernatant. Peripheral blood neutrophil, lymphocyte, and platelet counts decreased in all the virus-inoculated calves but were significantly lower and remained decreased longer in the calves given HV24515. For each isolate, a decrease in the number of mature myeloid cells in the bone marrow coincided with the development of neutropenia, but the depletion persisted significantly longer (4 to 6 d) in the calves given HV24515. In the bone marrow of calves given LV11Q, the number of proliferating myeloid cells increased in proportion to the decrease in the number of mature myeloid cells. In the calves inoculated with HV24515, BVDV antigen was observed in bone marrow cells when the peripheral blood counts were lowest. Megakaryocytes were the predominant cell type exhibiting positive BVDV staining; myeloid cells rarely stained positively. Viral antigen was not observed in the bone marrow of calves given LV11Q. These experiments demonstrated that ncpBVDV-2 isolates of both high and low virulence caused decreased leukocyte and platelet counts, but only the high-virulence HV24515 isolate caused a delay in the production of myeloid proliferating cells. The delay may contribute to the ability of certain ncpBVDV-2 isolates to induce severe disease. PMID:14979434

  17. Lipolytic enzymes in bovine thyroid tissue. I. Subcellular localization, purification and characterization of acid phospholipase A1.

    PubMed

    De Wolf, M; Lagrou, A; Hilderson, H J; Dierick, W

    1978-12-01

    In mammalian cells the catabolism of membrane phosphoglycerides proceeds probably entirely through a deacylation pathway catalysed by phospholipase A and lysophospholipase (Wise & Elwyn, 1965). In the initial attack of diacylphosphoglycerides by phospholipase A two enzymatic activities with different positional specificities have been distinguished: phospholipase A1 (phosphatidate 1-acyl hydrolase EN 3.1.1.32) and phospholipase A2 (phosphatidate 2-acyl hydrolase EN 3.1.1.4) (Van Deenen & De Haas, 1966). Studies on these intracellular phospholipases were mainly concerned with their subcellular localization. Only occasionally more detailed enzymatic investigations have been conducted on them, in contrast to export phospholipases e.g. from snake venom, bee venom and porcine pancreas, which have been extensively investigated (Brockerhoff & Jensen 1974a). In a previous paper (De Wolf et al., 1976a), the presence of phospholipase A1 and phospholipase A2 activities in bovine thyroid was demonstrated, using 1-[9, 10-3H] stearoyl-2-[1-14C] linoleyl-sn-glycero-3-phosphocholine as a substrate. Optimal activity was observed in both instances at pH 4. Addition of the anionic detergent sodium taurocholate increased the A2 type activity and decreased the A1 type activity suggesting the presence of different enzymes. The lack of influence of Ca2+-ions and EDTA and the acid pH optima could suggest lysosomal localization. In this paper the subcellular distribution of both acid phospholipase activities is described as well as a purification scheme for phospholipase A1. Some characteristics of the purified enzyme preparation are discussed.

  18. In vitro properties and performance of glutaraldehyde-crosslinked bovine pericardial bioprostheses treated with glutamic acid.

    PubMed

    Braile, Maria Christiane Valéria Braga; Carnevalli, Nelly Cristina; Goissis, Gilberto; Ramirez, Vladimir Aparecido; Braile, Domingo Marcolino

    2011-05-01

    Calcification is one of the major causes of failure of heart valve bioprostheses (HVBs) derived from glutaraldehyde (GA)-processed bovine pericardium (BP) or porcine aortic valves. New crosslinking reagent procedures are still far from giving satisfactory results, and this is the main reason why GA is still the reagent of choice for the fixation of native tissue intended for HVB manufacture. Nevertheless, two new findings with respect to GA processing may significantly improve HVB performance postimplantation: the finding that increasing concentrations of GA result in a decrease in calcification; the blocking of free aldehyde usually by nucleophyles or the treatment of processed material at low pH. This work investigates the in vitro properties of BP fixed with GA followed by the treatment with glutamic acid under alkaline conditions in order to prepare BP materials with lower calcification potential postimplantation. In comparison to conventional processing, except for the tensile strength that was slightly lower, elongation and toughness were higher than the accepted values. No significant differences were observed in the performance indexes (mean pressure gradient, mean effective area, regurgitant fraction, performance and efficiency indexes) with wear resistance over 150 × 10⁶ cycles. These results indicate that the processing of BP described in this work may be of potential use in the manufacture of HVBs.

  19. A Digital Gene Expression-Based Bovine Gene Atlas Evaluating 92 Adult, Juvenile and Fetal Cattle Tissues

    USDA-ARS?s Scientific Manuscript database

    A comprehensive transcriptome survey, or “Gene Atlas,” provides information essential for a complete understanding of the genomic biology of an organism. Using a digital gene expression approach, we developed a Gene Atlas of RNA abundance in 92 adult, juvenile and fetal cattle tissues. The samples...

  20. Monitoring quinolone antibacterial residues in bovine tissues: extraction with hot water and liquid chromatography coupled to a single- or triple-quadrupole mass spectrometer.

    PubMed

    Bogialli, Sara; D'Ascenzo, Giuseppe; Di Corcia, Antonio; Innocenti, Giorgiana; Laganà, Aldo; Pacchiarotta, Tiziana

    2007-01-01

    A rapid and sensitive procedure for determining residues of seven quinolone antibacterials in bovine muscle, kidney and liver is presented. The method is based on the matrix solid-phase dispersion (MSPD) technique with hot water as extractant followed by liquid chromatography/single quadrupole mass spectrometry (LC/MS) or triple-quadrupole mass spectrometry (LC/MS/MS). After dispersing tissue samples on hydrazine sulfate treated sand, target compounds were eluted from the MSPD column by passing through it 4 mL of water heated at 100 degrees C. After pH adjustment and filtration, 200 and 5 microL of the aqueous extracts were respectively injected into the LC/MS and LC/MS/MS instruments. With the former instrument, MS data were acquired in the three-ion selected ion monitoring mode, while MS/MS data acquisition was performed in the multi-reaction monitoring mode by selecting two precursor ion to product ion transitions for each target compound. Hot water appeared to be an efficient extracting medium, since absolute recoveries of the analytes were 84-102%. Using norfloxacin (a quinolone not used in veterinary medicine) as surrogate internal standard, the accuracy of the method at three concentration levels equal to 0.5, 1 and 1.5 times the maximum residue limits (MRLs) set by the european union was 88-109% with relative standard deviations (RSDs) not higher than 7%. The use of LC/MS/MS allowed detection and quantification of the analytes in any tissue considered to be performed at concentrations by far lower than half of their MRLs. Vice versa, the single-quadrupole MS arrangement, while succeeding in monitoring quinolones in muscle tissue at the 0.5 MRL level, showed to be not sufficiently selective for unambiguous identification of some quinolones in kidney and liver.

  1. Human urinary bladder regeneration through tissue engineering - an analysis of 131 clinical cases.

    PubMed

    Pokrywczynska, Marta; Adamowicz, Jan; Sharma, Arun K; Drewa, Tomasz

    2014-03-01

    Replacement of urinary bladder tissue with functional equivalents remains one of the most challenging problems of reconstructive urology over the last several decades. The gold standard treatment for urinary diversion after radical cystectomy is the ileal conduit or neobladder; however, this technique is associated with numerous complications including electrolyte imbalances, mucus production, and the potential for malignant transformation. Tissue engineering techniques provide the impetus to construct functional bladder substitutes de novo. Within this review, we have thoroughly perused the literature utilizing PubMed in order to identify clinical studies involving bladder reconstruction utilizing tissue engineering methodologies. The idea of urinary bladder regeneration through tissue engineering dates back to the 1950s. Many natural and synthetic biomaterials such as plastic mold, gelatin sponge, Japanese paper, preserved dog bladder, lyophilized human dura, bovine pericardium, small intestinal submucosa, bladder acellular matrix, or composite of collagen and polyglycolic acid were used for urinary bladder regeneration with a wide range of outcomes. Recent progress in the tissue engineering field suggest that in vitro engineered bladder wall substitutes may have expanded clinical applicability in near future but preclinical investigations on large animal models with defective bladders are necessary to optimize the methods of bladder reconstruction by tissue engineering in humans.

  2. Agonists of the G protein-coupled receptor 109A-mediated pathway promote antilipolysis by reducing serine residue 563 phosphorylation of hormone-sensitive lipase in bovine adipose tissue explants.

    PubMed

    Kenéz, A; Locher, L; Rehage, J; Dänicke, S; Huber, K

    2014-01-01

    A balanced lipolytic regulation in adipose tissues based on fine-tuning of prolipolytic and antilipolytic pathways is of vital importance to maintain the metabolic health in dairy cows. Antilipolytic pathways, such as the G protein-coupled receptor 109A (GPR109A)-mediated pathway and the insulin signaling pathway in bovine adipose tissues may be involved in prohibiting excessive lipomobilization by reducing triglycerol hydrolysis. This study aimed to evaluate the in vitro antilipolytic potential of the mentioned pathways in bovine adipose tissue explants. Therefore, subcutaneous and retroperitoneal adipose tissue samples (approximately 100mg) of German Holstein cows were treated for 90 min ex vivo with nicotinic acid (2, 8, or 32 μM), nicotinamide (2, 8, or 32 μM), β-hydroxybutyrate (0.2, 1, or 5mM), or insulin (12 mU/L), with a concurrent lipolytic challenge provoked with 1 μM isoproterenol. Lipolytic and antilipolytic responses of the adipose tissues were assessed by measuring free glycerol and nonesterified fatty acid release. To identify molecular components of the investigated antilipolytic pathways, protein abundance of GPR109A and the extent of hormone-sensitive lipase (HSL) phosphorylation at serine residue 563 were detected by Western blotting. Treatment with nicotinic acid or β-hydroxybutyrate decreased the lipolytic response in adipose tissue explants and concurrently reduced the extent of HSL phosphorylation, but treatment with nicotinamide or insulin did not. Subcutaneous adipose tissue constitutively expressed more GPR109A protein, but no other depot-specific differences were observed. This study provides evidence that the GPR109A-mediated pathway is functionally existent in bovine adipose tissues, and confirms that HSL phosphorylation at serine residue 563 is also important in antilipolytic regulation in vitro. This antilipolytic pathway may be involved in a balanced lipid mobilization in the dairy cow. Copyright © 2014 American Dairy Science

  3. Effect of dietary fat on uptake of lysine, phenylalanine, leucine and methionine by bovine mammary tissue slices in vitro

    SciTech Connect

    Nianogo, A.J.; Amos, H.E.; Dean, R.; Froetschel, A. ); Fernandez, J.M. )

    1989-08-01

    Four mature Holstein cows in late lactation were blocked in two groups based on milk production, in a 2x2 reversal with 21-day periods, and fed: (A) control diet; (B) A plus 1 kg/day tallow. Cows were fed sorghum silage ad libitum. Blood samples were collected from the jugular vein on day 15, 17, and 19 of each period. Fat did not effect DM intake or milk yield, however milk CP yield was 20% lower. Plasma lipids increased 33.6%, glucose decreased 9% and insulin/glucagon ratio decreased 21.2% in cow fed fat. After period two, cows were slaughtered and mammary tissue sampled for incubation in Krebs Ringer bicarbonate buffer containing 22 AA at arterial concentration and .225 {mu}Ci/ml of {sup 14}C-labelled L-Leu, L-Phe, L-Lys or D/L Met. Dietary fat decreased tissue AA uptake rate by 21.2%. Uptake was 4.8, 10.3, 17.8 and 2.4 {times} 10{sup {minus}3} {mu}M/min/gm of tissue DM for Phe, Lys, Leu and Met, respectively. Results suggest that dietary fat may decrease milk protein synthesis by lowering the rate of AA uptake.

  4. Tissue/fluid correlation study for the depletion of sulfadimethoxine in bovine kidney, liver, plasma, urine, and oral fluid.

    PubMed

    Chiesa, O A; Li, H; Kijak, P J; Li, J X; Lancaster, V; Smith, M L; Heller, D N; Thomas, M H; Von Bredow, J

    2012-06-01

    Sulfonamides are among the oldest, but still effective, antimicrobial veterinary medicines. In steers and dairy cows, the sulfonamides are effective in the treatment of respiratory disease and general infections. Sulfadimethoxine (SDM) has been approved by US Food and Drug Administration (FDA) for use in steers and dairy cows with a tolerance of 100 ng/g (ppb) in edible tissues and 10 ppb in milk. The detection of SDM residue above tolerance in the animal slaughtered for food process will result in the whole carcass being discarded. This report describes a comprehensive depletion study of SDM (and its main metabolite) in plasma, urine, oral fluid, kidney, and liver. In this study, nine steers were injected intravenously with the approved dose of SDM; the loading dose was 55 mg/kg, followed by 27.5 mg/kg dose at 24 h and again at 48 h. Fluids (blood, urine, and saliva) and tissue (liver and kidney) samples were collected at intervals after the last dose of SMD. The combination of laparoscopic serial sampling technique with the liquid chromatography/mass spectrometry method provided the data to establish the tissue/fluid correlation in the depletion of SMD. A strong correlation and linearity of the log-scale concentration over time in the depletion stage has been confirmed for kidney, liver, and plasma.

  5. Automated pericardium delineation and epicardial fat volume quantification from noncontrast CT.

    PubMed

    Ding, Xiaowei; Terzopoulos, Demetri; Diaz-Zamudio, Mariana; Berman, Daniel S; Slomka, Piotr J; Dey, Damini

    2015-09-01

    The authors aimed to develop and validate an automated algorithm for epicardial fat volume (EFV) quantification from noncontrast CT. The authors developed a hybrid algorithm based on initial segmentation with a multiple-patient CT atlas, followed by automated pericardium delineation using geodesic active contours. A coregistered segmented CT atlas was created from manually segmented CT data and stored offline. The heart and pericardium in test CT data are first initialized by image registration to the CT atlas. The pericardium is then detected by a knowledge-based algorithm, which extracts only the membrane representing the pericardium. From its initial atlas position, the pericardium is modeled by geodesic active contours, which iteratively deform and lock onto the detected pericardium. EFV is automatically computed using standard fat attenuation range. The authors applied their algorithm on 50 patients undergoing routine coronary calcium assessment by CT. Measurement time was 60 s per-patient. EFV quantified by the algorithm (83.60 ± 32.89 cm(3)) and expert readers (81.85 ± 34.28 cm(3)) showed excellent correlation (r = 0.97, p < 0.0001), with no significant differences by comparison of individual data points (p = 0.15). Voxel overlap by Dice coefficient between the algorithm and expert readers was 0.92 (range 0.88-0.95). The mean surface distance and Hausdorff distance in millimeter between manually drawn contours and the automatically obtained contours were 0.6 ± 0.9 mm and 3.9 ± 1.7 mm, respectively. Mean difference between the algorithm and experts was 9.7% ± 7.4%, similar to interobserver variability between 2 readers (8.0% ± 5.3%, p = 0.3). The authors' novel automated method based on atlas-initialized active contours accurately and rapidly quantifies EFV from noncontrast CT.

  6. Evaluation of tissue-engineered bone constructs using rabbit fetal osteoblasts on acellular bovine cancellous bone matrix

    PubMed Central

    Rashmi; Pathak, Rekha; Amarpal; Aithal, H. P.; Kinjavdekar, P.; Pawde, A. M.; Tiwari, A. K.; Sangeetha, P.; Tamilmahan, P.; Manzoor, A. B.

    2017-01-01

    Aim: The aim of this study was to generate composite bone graft and investigate the rabbit fetal osteoblasts adhesion, proliferation and penetration on acellular matrices of cancellous bone. Materials and Methods: Acellular cancellous bone was prepared and developed as in the previous study with little modification. These matrices were decellularized by rapid freeze and thaw cycle. To remove the cell debris, they were then treated with hydrogen peroxide (3%) and ethanol to remove antigenic cellular and nuclear materials from the scaffold. Primary osteoblast cells were harvested from 20 to 22 days old rabbit fetal long and calvarial bone. These cells were cultured and characterized using a specific marker. The third passaged fetal osteoblast cells were then seeded on the scaffold and incubated for 14 days. The growth pattern of the cells was observed. Scanning electron microscope and hematoxylin and eosin staining were used to investigate cells proliferation. Results: The cells were found to be growing well on the surface of the scaffold and were also present in good numbers with the matrix filopodial extensions upto inside of the core of the tissue. Conclusion: Thus, a viable composite scaffold of bone could be developed which has a great potential in the field of bone tissue engineering. PMID:28344398

  7. Differential effects of propionate or β-hydroxybutyrate on genes related to energy balance and insulin sensitivity in bovine white adipose tissue explants from a subcutaneous and a visceral depot.

    PubMed

    Hosseini, A; Behrendt, C; Regenhard, P; Sauerwein, H; Mielenz, M

    2012-08-01

    Ruminants rely on short-chain fatty acids (SCFA) as principal energy source. Herein, we compared the effects of propionate, β-hydroxybutyrate (BHB) and insulin on mRNA abundance of energy balance-related genes by short-term incubation (4 h) in bovine subcutaneous (SC) and retroperitoneal (RP) adipose tissue (AT) explants in vitro. Propionate either significantly (p < 0.05), or as a trend (p ≤ 0.1) affected mRNA abundance of genes such as adiponectin system in both depots in treated samples versus controls. Propionate increased adiponectin receptor 1 (AdipoR1) and AdipoR2 mRNA only in SC AT. β-hydroxybutyrate decreased mRNA abundance of adiponectin and AdipoR1 in SC AT as a trend. The mRNA abundance of free fatty acid receptor 2/3 (FFAR2/3) and other genes of interest (GOI) was increased during differentiation in bovine preadipocyte culture. The mRNA abundance of all the GOI remained unchanged after short-term insulin stimulation. In total, propionate, BHB or insulin during short-term treatment exert divergent effects on the mRNA abundance of GOI in both depots in vitro. Our results indicate that the bovine adiponectin system might be more sensitive to propionate than to BHB. We demonstrated the presence of FFAR2/3 mRNA not only in both AT depots but also in differentiating preadipocytes isolated from bovine SC AT. Therefore, we established that SCFA are able to exert insulin-independent effects on bovine adipose tissue, which might be independent from propionate uptake-related events. © 2011 Blackwell Verlag GmbH.

  8. [Diseases of the pericardium in Avenzoar's Kitab al-Taysir (c.1095-1162)].

    PubMed

    Pena, C; Giron, F; Moreno, R M

    1997-01-01

    Avenzoar has been credited as the author of the first description of inflammation of the pericardium in medical historical literature. Our study shows that although Avenzoar authored a study of diseases of the pericardium with emphasis on pathologies, his epistemological framework was similar to that used by Galen and Avicenna, authors who constituted the source of knowledge for Islamic medicine. We show that the approach used by Avenzoar appears to derive from the absence of anatomical and physiological information, and from a detailed description of the indications and treatments, which distinguish his work from earlier writings.

  9. Seaweed floating in the pericardium: a rare case of primary dedifferentiated liposarcoma.

    PubMed

    Zanini, Gregoriana; Gorga, Elio; Pasini, Federica; Salemme, Marianna; Petrilli, Giulia; Bercich, Luisa; Lorusso, Roberto; Pasini, Gian Franco

    2016-01-01

    Primary cardiac tumors are uncommon and primary liposarcoma of the pericardium is extremely rare. We describe the case of a 55-year-old Caucasian woman without significant medical history, who presented with 3weeks complain of dyspnea, peripheral edema, and gain weight. Echocardiography revealed a huge pericardial mass confirmed by computed tomography and by magnetic resonance. The lesion was primitive of the pericardium but the surgery was not able to cut it off because of the absence of cleavage planes. Histopathologic analysis detected a dedifferentiated liposarcoma. Mediastinum compression syndrome brought the patient to the exitus in a few days. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Congenital partial defect of the left pericardium presenting as chest pain.

    PubMed

    Jeong, Heui-Jeong; Seol, Sang-Hoon; Seo, Guang-Won; No, Tae-Hoon; Seo, Min-Gyo; Park, Bo-Min; Song, Pil-Sang; Kim, Dong-Kie; Kim, Ki-Hun; Kim, Doo-Il

    2014-12-01

    Congenital pericardial defect is a rare cardiac defect with variable clinical presentations. It is usually an unexpected finding during cardiac surgery or autopsy. The clinical detection of congenital absence of pericardium is important because of its life-threatening complications such as fatal myocardial strangulation, myocardial ischaemia and sudden death. We present a patient with the incidental finding of left-sided partial defect of the pericardium during evaluation of chest pain. Copyright © 2014 Australian and New Zealand Society of Cardiac and Thoracic Surgeons (ANZSCTS) and the Cardiac Society of Australia and New Zealand (CSANZ). Published by Elsevier B.V. All rights reserved.

  11. Symptomatic Partial Congenital Absence of the Pericardium Revealed Using Cardiac Magnetic Resonance.

    PubMed

    Valero, Ernesto; Ferrero, Jose Antonio; López-Lereu, Maria Pilar; Chorro, Francisco Javier

    2015-10-01

    Congenital absence of the pericardium is a very uncommon finding, and its diagnosis poses a challenge because it is rarely suspected in daily clinical practice. Although in most cases it has a benign course, this congenital defect should be identified because of the associated risk of sudden death. We present a symptomatic case of partial congenital absence of the left pericardium suspected as the result of an abnormal response to exercise stress testing, and confirmed using cardiac magnetic resonance imaging. We review the current diagnostic tools and therapeutic indications of this rare anomaly. Copyright © 2015 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.

  12. Plasma leptin and mRNA expression of lipogenesis and lipolysis-related factors in bovine adipose tissue around parturition.

    PubMed

    Sadri, H; Mielenz, M; Morel, I; Bruckmaier, R M; van Dorland, H A

    2011-12-01

    The objective was to study changes in plasma leptin concentration parallel to changes in the gene expression of lipogenic- and lipolytic-related genes in adipose tissue of dairy cows around parturition. Subcutaneous fat biopsies were taken from 27 dairy cows in week 8 antepartum (a.p.), on day 1 postpartum (p.p.) and in week 5 p.p. Blood samples were assayed for concentrations of leptin and non-esterified fatty acids (NEFA). Subcutaneous adipose tissue was analysed for mRNA abundance by real-time qRT-PCR encoding for leptin, adiponectin receptor 1 (AdipoR1), adiponectin receptor 2 (AdipoR2), hormones-sensitive lipase (HSL), perilipin (PLIN), lipoprotein lipase (LPL), acyl-CoA synthase long-chain family member 1 (ACSL1), acetyl-CoA carboxylase (ACC), fatty acid synthase (FASN) and glycerol-3-phosphate dehydrogenase 2 (GPD2). Body weight and body condition score of the cows were lower after parturition than before parturition. The calculated energy balance was negative in week 1 and 5 p.p., with higher negative energy balance in week 1 p.p. compared with that in week 5 p.p. On day 1 p.p., highest concentrations of NEFA (353.3 μmol/l) were detected compared with the other biopsy time-points (210.6 and 107.7 μmol/l, in week 8 a.p., and week 5 p.p. respectively). Reduced plasma concentrations of leptin during p.p. when compared with a.p. would favour increasing metabolic efficiency and energy conservation for mammary function and reconstitution of body reserves. Lower mRNA abundance of ACC and FASN expression on day 1 p.p. compared with other biopsy time-points suggests an attenuation of fatty acid synthesis in subcutaneous adipose tissue shortly after parturition. Gene expression of AdipoR1, AdipoR2, HSL, PLIN, LPL, ACSL1 and GPD2 was unchanged over time.

  13. Effects of Alcohol on Postoperative Adhesion Formation in Ischemic Myocardium and Pericardium.

    PubMed

    Elmadhun, Nassrene Y; Sabe, Ashraf A; Lassaletta, Antonio D; Dalal, Rahul S; Sellke, Frank W

    2017-08-01

    Postoperative formation of adhesions increases risk of complications during cardiac reoperations. We previously demonstrated that swine supplemented with vodka had a significant reduction in adhesions at sternotomy after previous thoracotomy. This follow-up study was conducted to determine reproducibility and the mechanism for adhesion reduction in swine supplemented with ethanol. An ameroid constrictor was placed in the left circumflex in 14 male Yorkshire swine to induce chronic myocardial ischemia through left minithoracotomy. Animals were supplemented postoperatively with ethanol (45 g ETOH, n = 7) or sucrose (80 g SUC, n = 7) for 7 weeks, followed by a reoperative median sternotomy. The ETOH group had significantly fewer adhesions, thinner pericardial thickness, decreased intramyocardial fibrosis, and decreased myocardial collagen deposition compared with the SUC group. In the myocardium, ETOH animals had decreased expression of proadhesion proteins focal adhesion kinase, paxillin, integrin-β1, transforming growth factor-β1 and phosphorylated SMAD and increased expression of adhesion breakdown proteins matrix metalloproteinase (MMP) 1, MMP2, MMP3, and MMP9 compared with SUC animals. However in the pericardium, ETOH animals had increased expression of proadhesion proteins focal adhesion kinase, paxillin, phosphorylated paxillin, vinculin, integrin-β1, tumor necrosis factor-α, transforming growth factor-β, and phosphorylated SMAD3, and decreased expression of adhesion breakdown proteins MMP1, MMP3, MMP9, and plasmin compared with SUC animals. Alcohol supplementation substantially reduced postoperative pericardial adhesion formation, attenuated pericardial thickening, and reduced myocardial fibrosis in response to chronic ischemia. Alcohol supplementation modulates adhesion protein and MMP/tissue inhibitor of metalloproteinase expression, favoring a profile associated with reduced pericardial adhesions. These results suggest that the myocardium is the

  14. Multipotency and cardiomyogenic potential of human adipose-derived stem cells from epicardium, pericardium, and omentum.

    PubMed

    Wystrychowski, Wojciech; Patlolla, Bhagat; Zhuge, Yan; Neofytou, Evgenios; Robbins, Robert C; Beygui, Ramin E

    2016-06-13

    Acute myocardial infarction (MI) leads to an irreversible loss of proper cardiac function. Application of stem cell therapy is an attractive option for MI treatment. Adipose tissue has proven to serve as a rich source of stem cells (ADSCs). Taking into account the different morphogenesis, anatomy, and physiology of adipose tissue, we hypothesized that ADSCs from different adipose tissue depots may exert a diverse multipotency and cardiogenic potential. The omental, pericardial, and epicardial adipose tissue samples were obtained from organ donors and patients undergoing heart transplantation at our institution. Human foreskin fibroblasts were used as the control group. Isolated ADSCs were analyzed for adipogenic and osteogenic differentiation capacity and proliferation potential. The immunophenotype and constitutive gene expression of alkaline phosphatase (ALP), GATA4, Nanog, and OCT4 were analyzed. DNA methylation inhibitor 5-azacytidine was exposed to the cells to stimulate the cardiogenesis. Finally, reprogramming towards cardiomyocytes was initiated with exogenous overexpression of seven transcription factors (ESRRG, GATA4, MEF2C, MESP1, MYOCD, TBX5, ZFPM2) previously applied successfully for fibroblast transdifferentiation toward cardiomyocytes. Expression of cardiac troponin T (cTNT) and alpha-actinin (Actn2) was analyzed 3 weeks after initiation of the cardiac differentiation. The multipotent properties of isolated plastic adherent cells were confirmed with expression of CD29, CD44, CD90, and CD105, as well as successful differentiation toward adipocytes and osteocytes; with the highest osteogenic and adipogenic potential for the epicardial and omental ADSCs, respectively. Epicardial ADSCs demonstrated a lower doubling time as compared with the pericardium and omentum-derived cells. Furthermore, epicardial ADSCs revealed higher constitutive expression of ALP and GATA4. Increased Actn2 and cTNT expression was observed after the transduction of seven

  15. The Resistance of Electron Transport Chain Fe-S Clusters to Oxidative Damage during the Reaction of Peroxynitrite with Mitochondrial Complex II and Rat Heart Pericardium

    PubMed Central

    Pearce, Linda L.; Martinez-Bosch, Sandra; Manzano, Elisenda Lopez; Winnica, Daniel E.; Epperly, Michael W.; Peterson, Jim

    2009-01-01

    The effects of peroxynitrite and nitric oxide on the iron-sulfur clusters in complex II (succinate dehydrogenase) isolated from bovine heart have been studied primarily by EPR spectroscopy and no measurable damage to the constitutive 2Fe-2S, 3Fe-4S, or 4Fe-4S clusters was observed. The enzyme can be repeatedly oxidized with a slight excess of peroxynitrite and then quantitatively re-reduced with succinate. When added in large excess, peroxynitrite reacted with at least one tyrosine in each subunit of complex II to form 3-nitrotyrosines, but activity was barely compromised. Examination of rat-heart pericardium subjected to conditions leading to peroxynitrite production showed a small inhibition of complex II (16%) and a greater inhibition of aconitase (77%). In addition, experiments performed with excesses of sodium citrate and sodium succinate on rat-heart pericardium indicated that the “g ~2.01” EPR signal observed immediately following the beginning of conditions modeling oxidative/nitrosative stress, could be a consequence of both reversible oxidation of the constitutive 3Fe-4S cluster in complex II and degradation of the 4Fe-4S cluster in aconitase. However, the net signal envelope, which becomes apparent in less than one minute following the start of oxidative/nitrosative conditions, is dominated by the component arising from complex II. Taking into account the findings of a previous study concerning complexes I and III [L.L. Pearce, A.J. Kanai, M.W. Epperly, J. Peterson (2005) Nitric Oxide 13, 254-63] it is now apparent that, with the exception of the cofactor in aconitase, mammalian (mitochondrial) iron-sulfur clusters are surprisingly resistant to degradation stemming from oxidative/nitrosative stress. PMID:19118636

  16. [Clinical case of the month. Constrictive pericarditis with a macroscopically normal pericardium: apropos of a case].

    PubMed

    Hoffer, E; Materne, P; Limet, R; Boland, J

    2007-04-01

    The diagnosis of constrictive pericarditis is not easy to make. This rare condition can be suggested by clinical, echocardiograohic, hemodynamic, and radiological signs. It must be distinguished from restrictive cardiomyopathy as therapeutic options are radically different. We present an ambiguous case of constrictive pericarditis with macroscopically normal pericardium recognized 10 years after open-chest cardiac surgery: a large pericardiectomy rapidly induced clinical improvement.

  17. Pericardium Plug in the Repair of the Corneoscleral Fistula After Ahmed Glaucoma Valve Explantation

    PubMed Central

    Yoo, Chungkwon; Kwon, Sung Wook

    2008-01-01

    We report four cases in which a pericardium (Tutoplast®) plug was used to repair a corneoscleral fistula after Ahmed Glaucoma Valve (AGV) explantation. In four cases in which the AGV tube had been exposed, AGV explantation was performed using a pericardium (Tutoplast®) plug to seal the defect previously occupied by the tube. After debridement of the fistula, a piece of processed pericardium (Tutoplast®), measured 1 mm in width, was plugged into the fistula and secured with two interrupted 10-0 nylon sutures. To control intraocular pressure, a new AGV was implanted elsewhere in case 1, phaco-trabeculectomy was performed concurrently in case 2, cyclophotocoagulation was performed postoperatively in case 3 and anti-glaucomatous medication was added in case 4. No complication related to the fistula developed at the latest follow-up (range: 12~26 months). The pericardium (Tutoplast®) plug seems to be an effective method in the repair of corneoscleral fistulas resulting from explantation of glaucoma drainage implants. PMID:19096247

  18. Use of equine pericardium sheet (LYOMESH®) as dura mater substitute in endoscopic endonasal transsphenoidal surgery

    PubMed Central

    Cavallo, Luigi M.; Solari, Domenico; Somma, Teresa; Di Somma, Alberto; Chiaramonte, Carmela; Cappabianca, Paolo

    Objective The aim of this study was to describe the use of equine pericardium sheet (Lyomesh ® ) as dural substitute for sellar reconstruction after endoscopic endonasal transsphenoidal surgery for the removal of pituitary adenomas. Methods We reviewed data of patients that underwent surgery by means of an endoscopic endonasal transsphenoidal approach for the removal of pituitary adenomas over a 12-months period, starting in May 2012, i.e. when we adopted Lyomesh ® (Audio Technologies, Piacenza, Italy) an equine pericardium sheet, as dura mater substitute. Results: During the 12-months period evaluated, we performed an endoscopic endonasal transsphenoidal operation for a variety of pituitary lesions on 102 consecutive patients. Among these, in 12 patients (9.4%) harboring a pituitary adenoma, the implant of the pericardium sheet was used. Four patients (33.3%) presented a small intraoperative cerebrospinal fluid (CSF) leak; in these cases the Lyomesh ® was placed intradurally with fibrin glue and, thereafter, several layers were positioned in extradural space. In 8 other subjects without any evidence of CSF leak, the dural substitute was placed intradurally and fibrin glue was injected intradurally to hold the material in place. Conclusions: Even if based on a relatively small patient series, our experience demonstrated that the use of equine pericardium sheet (Lyomesh ® ) as dura mater substitute in transsphenoidal surgery is safe and biocompatible, as compared with other dural substitutes. PMID:24251248

  19. Back-scattered electron imaging and elemental analysis of retrieved bone tissue following sinus augmentation with deproteinized bovine bone or biphasic calcium phosphate.

    PubMed

    Lindgren, Christer; Hallman, Mats; Sennerby, Lars; Sammons, Rachel

    2010-09-01

    To compare resorption of a synthetic biphasic calcium phosphate (BCP) bone-graft substitute with deproteinized bovine bone (DBB) used for human maxillary sinus augmentation. Eleven patients underwent bilateral maxillary sinus floor augmentation with DBB in one side and a BCP (40%beta-tricalcium phosphate (beta-TCP) and 60% hydroxyapatite) in the contralateral side. Simultaneously, with the augmentation on each side a microimplant was placed vertically from the top of the alveolar crest penetrating the residual bone and the grafting material. Eight months after initial surgery the microimplants were retrieved with a surrounding bone core. The composition of residual graft material and surrounding bone was analysed by scanning electron microscopy and energy dispersive X-ray spectroscopy. Residual graft material of both types was present as 10-500 mum particles in direct contact with, or completely surrounded by, newly formed bone; smaller particles were also present in non-mineralized tissue. In the case of BCP the bone-graft substitute interface showed evidence of superficial disintegration of particles into individual grains. Median Ca/P ratios (at.%), determined from >200 discreet sites within residual graft particles and adjacent bone, were: DBB: 1.61 (confidence interval [CI] 1.59-1.64); BCP: 1.5 (CI 1.45-1.52); DBB-augmented bone: 1.62 (CI 1.59-1.66); BCP-augmented bone: 1.52 (CI 1.47-1.55); P=0.028 for DBB vs. BCP and DBB- vs. BCP-augmented bone. The reduction in Ca/P ratio for BCP over the healing period is consistent with the dissolution of beta-TCP and reprecipitation on the surface of calcium-deficient hydroxyapatite. The beta-TCP component of BCP may be gradually substituted by calcium-deficient hydroxyapatite over the healing period. This process and superficial degranulation of BCP particles may influence the progress of resorption and healing.

  20. Canine Platelet Lysate Is Inferior to Fetal Bovine Serum for the Isolation and Propagation of Canine Adipose Tissue- and Bone Marrow-Derived Mesenchymal Stromal Cells

    PubMed Central

    Russell, Keith A.; Gibson, Thomas W. G.; Chong, Andrew; Co, Carmon; Koch, Thomas G.

    2015-01-01

    Background Mesenchymal stromal cells (MSC) are increasingly investigated for their clinical utility in dogs. Fetal bovine serum (FBS) is a common culture supplement used for canine MSC expansion. However, FBS content is variable, its clinical use carries risk of an immune response, and its cost is increasing due to global demand. Platelet lysate (PL) has proven to be a suitable alternative to FBS for expansion of human MSC. Hypothesis and Objectives We hypothesized that canine adipose tissue (AT) and bone marrow (BM) MSC could be isolated and expanded equally in PL and FBS at conventionally-used concentrations with differentiation of these MSC unaffected by choice of supplement. Our objectives were to evaluate the use of canine PL in comparison with FBS at four stages: 1) isolation, 2) proliferation, 3) spontaneous differentiation, and 4) directed differentiation. Results 1) Medium with 10% PL was unable to isolate MSC. 2) MSC, initially isolated in FBS-supplemented media, followed a dose-dependent response with no significant difference between PL and FBS cultures at up to 20% (AT) or 30% (BM) enrichment. Beyond these respective peaks, proliferation fell in PL cultures only, while a continued dose-dependent proliferation response was noted in FBS cultures. 3) Further investigation indicated PL expansion culture was inducing spontaneous adipogenesis in concentrations as low as 10% and as early as 4 days in culture. 4) MSC isolated in FBS, but expanded in either FBS or PL, maintained ability to undergo directed adipogenesis and osteogenesis, but not chondrogenesis. Conclusions/Significance Canine PL did not support establishment of MSC colonies from AT and BM, nor expansion of MSC, which appear to undergo spontaneous adipogenesis in response to PL exposure. In vivo studies are warranted to determine if concurrent use of MSC with any platelet-derived products such as platelet-rich plasma are associated with synergistic, neutral or antagonistic effects. PMID:26353112

  1. Actions of activin A, connective tissue growth factor, hepatocyte growth factor and teratocarcinoma-derived growth factor 1 on the development of the bovine preimplantation embryo.

    PubMed

    Kannampuzha-Francis, Jasmine; Tribulo, Paula; Hansen, Peter J

    2016-05-17

    The reproductive tract secretes bioactive molecules collectively known as embryokines that can regulate embryonic growth and development. In the present study we tested four growth factors expressed in the endometrium for their ability to modify the development of the bovine embryo to the blastocyst stage and alter the expression of genes found to be upregulated (bone morphogenetic protein 15 (BMP15) and keratin 8, type II (KRT8)) or downregulated (NADH dehydrogenase 1 (ND1) and S100 calcium binding protein A10 (S100A10)) in embryos competent to develop to term. Zygotes were treated at Day 5 with 0.01, 0.1 or 1.0 nM growth factor. The highest concentration of activin A increased the percentage of putative zygotes that developed to the blastocyst stage. Connective tissue growth factor (CTGF) increased the number of cells in the inner cell mass (ICM), decreased the trophectoderm : ICM ratio and increased blastocyst expression of KRT8 and ND1. The lowest concentration of hepatocyte growth factor (HGF) reduced the percentage of putative zygotes becoming blastocysts. Teratocarcinoma-derived growth factor 1 increased total cell number at 0.01 nM and expression of S100A10 at 1.0 nM, but otherwise had no effects. Results confirm the prodevelopmental actions of activin A and indicate that CTGF may also function as an embryokine by regulating the number of ICM cells in the blastocyst and altering gene expression. Low concentrations of HGF were inhibitory to development.

  2. The use of bovine serum albumin-glutaraldehyde tissue adhesive (BioGlue(®)) for tumor bed closure following open partial nephrectomy.

    PubMed

    Bahouth, Z; Halachmi, S; Shprits, S; Burbara, Y; Avitan, O; Masarwa, I; Moskovitz, B; Nativ, O

    2017-10-01

    To report the results of the use of Bovine Serum Albumin-Glutaraldehyde tissue adhesive (BioGlue®) for tumor bed closure in open nephron-sparing surgery (NSS). The cohort included 255 patients with enhancing renal mass who underwent open NSS. We used open flank approach, with in-situ hypothermia and enucleation of the tumor. For tumor bed closure, we used the BioGlue(®) sealant for tumor bed filling, without suturing the edges. Mean patients' age was 65.4 years. 5.1% of patients had pre-operative chronic renal failure. Mean renal mass diameter was 4.2±1.6cm and mean R.E.N.A.L nephrometry score was 8.0±1.6. Mean ischemia time was 21.8±7.6. Mean estimated blood loss was 42±82ml and only two patients required blood transfusion. Urine leak and pseudo-aneurysm were recorded in two and one patient, respectively. None of the operations were converted to radical nephrectomy. The average change between post-operative and pre-operative eGFR (Δ=-1.7ml/min) was insignificant in a mean follow-up of 30.1±29.6 months. The 10-year recurrence-free survival rate was 99% and the 10-year overall survival rate was 85%. The use of BioGlue(®) alone for hemostasis after NSS is a feasible and safe alternative to classical suturing. Its use enables satisfactory functional outcome and could potentially reduce ischemia time. Copyright © 2017 AEU. Publicado por Elsevier España, S.L.U. All rights reserved.

  3. Validated method for determination of ultra-trace closantel residues in bovine tissues and milk by solid-phase extraction and liquid chromatography-electrospray ionization-tandem mass spectrometry.

    PubMed

    Sun, Hanwen; Wang, Fengchi; Ai, Lianfeng

    2007-12-21

    A liquid chromatographic-electrospray ionisation-tandem mass spectrometry method (LC-ESI-MS/MS) with solid extraction was developed and validated for the detection and determination of closantel residues in bovine tissues and milk. An acetonitrile-acetone mixture (80:20, v/v) was used for one-stage extraction of closantel residues in bovine tissues and milk samples, and the extract was cleaned by solid phase extraction with Oasis MAX cartridges. The mass spectrometer was operated in multiple reactions monitoring mode with negative electrospray interface. The limits of detection in different matrices were in the range of 0.008-0.009 microg/kg. The overall recoveries for bovine muscle, liver, kidney and milk samples spiked at four levels including MRL were in the range of 76.0-94.3%. The overall relative standard deviations were in the range of 3.57-8.61%. The linearity is satisfactory with a correlation coefficient (r(2)) of 0.9913-0.9987 at both concentration ranges of 0.02-100 microg/kg and 200-5000 microg/kg. The method is capable of identifying closantel residues at > or =0.02 microg/kg levels and was applied in the determination of closantel residues in animal origin foods.

  4. Slit-roundabout signaling regulates the development of the cardiac systemic venous return and pericardium.

    PubMed

    Mommersteeg, Mathilda T M; Andrews, William D; Ypsilanti, Athena R; Zelina, Pavol; Yeh, Mason L; Norden, Julia; Kispert, Andreas; Chédotal, Alain; Christoffels, Vincent M; Parnavelas, John G

    2013-02-01

    The Slit-Roundabout (Robo) signaling pathway has pleiotropic functions during Drosophila heart development. However, its role in mammalian heart development is largely unknown. To analyze the role of Slit-Robo signaling in the formation of the pericardium and the systemic venous return in the murine heart. Expression of genes encoding Robo1 and Robo2 receptors and their ligands Slit2 and Slit3 was found in or around the systemic venous return and pericardium during development. Analysis of embryos lacking Robo1 revealed partial absence of the pericardium, whereas Robo1/2 double mutants additionally showed severely reduced sinus horn myocardium, hypoplastic caval veins, and a persistent left inferior caval vein. Mice lacking Slit3 recapitulated the defects in the myocardialization, alignment, and morphology of the caval veins. Ligand binding assays confirmed Slit3 as the preferred ligand for the Robo1 receptor, whereas Slit2 showed preference for Robo2. Sinus node development was mostly unaffected in all mutants. In addition, we show absence of cross-regulation with previously identified regulators Tbx18 and Wt1. We provide evidence that pericardial defects are created by abnormal localization of the caval veins combined with ectopic pericardial cavity formation. Local increase in neural crest cell death and impaired neural crest adhesive and migratory properties underlie the ectopic pericardium formation. A novel Slit-Robo signaling pathway is involved in the development of the pericardium, the sinus horn myocardium, and the alignment of the caval veins. Reduced Slit3 binding in the absence of Robo1, causing impaired cardiac neural crest survival, adhesion, and migration, underlies the pericardial defects.

  5. Mid-term results of mitral valve repair with autologous pericardium in pediatric patients.

    PubMed

    Takahashi, Hiroaki; Kadowaki, Tasuku; Maruo, Ayako; Yutaka, Okita; Oshima, Yoshihiro

    2014-05-01

    Mitral valve repair to treat mitral regurgitation (MR) in pediatric patients remains challenging because of the complex morphology and fragility of the leaflets. The study aim was to review retrospectively the authors' experience with mitral valve repair using autologous pericardium. Between April 2004 and November 2011, nine pediatric patients (six males, three females; mean age 2.4 +/- 4.5 years) underwent mitral valve repair with an autologous fresh (n = 5) or glutaraldehyde-treated pericardium (n = 4) to treat severe MR. The etiology of the MR was acute endocarditis, acute chordal rupture and congenital disease in three, two, and four patients, respectively. Autologous pericardium was used for valvuloplasty, leaflet extension plasty and commissuroplasty in two, five, and two patients, respectively. Artificial chordal replacement was performed in three patients. No operative deaths or postoperative endocarditis occurred. One patient required mitral valve replacement at three days after the initial mitral repair. The most recent echocardiography findings of the remaining patients after a mean of 6.3 years (range: 1.3-9.0 years) showed that the severity of mitral insufficiency, left ventricular diastolic diameter and fractional shortening were 1.8 +/- 0.6 (grades 0-4), 40.4 +/- 8.4 mm (114.2 +/- 15.8% of normal) and 35.0 +/- 5.0%, respectively. Mitral valve repair using autologous fresh pericardium is associated with mid-term durability and resistance to infection when used for mitral valve repair of active endocarditis in pediatric patients. Augmentation with autologous pericardium could become an alternative to current surgical options.

  6. ProteINSIDE to Easily Investigate Proteomics Data from Ruminants: Application to Mine Proteome of Adipose and Muscle Tissues in Bovine Foetuses

    PubMed Central

    Kaspric, Nicolas; Picard, Brigitte; Reichstadt, Matthieu; Tournayre, Jérémy; Bonnet, Muriel

    2015-01-01

    Genomics experiments are widely acknowledged to produce a huge amount of data to be analysed. The challenge is to extract meaningful biological context for proteins or genes which is currently difficult because of the lack of an integrative workflow that hinders the efficiency and the robustness of data mining performed by biologists working on ruminants. Thus, we designed ProteINSIDE, a free web service (www.proteinside.org) that (I) provides an overview of the biological information stored in public databases or provided by annotations according to the Gene Ontology, (II) predicts proteins that are secreted to search for proteins that mediate signalisation between cells or tissues, and (III) analyses protein-protein interactions to identify proteins contributing to a process or to visualize functional pathways. Using lists of proteins or genes as a unique input, ProteINSIDE is an original all-in-one tool that merges data from these searches to present a fast overview and integrative analysis of genomic and proteomic data from Bovine, Ovine, Caprine, Human, Rat, and Murine species. ProteINSIDE was bench tested with 1000 proteins identifiers from each species by comparison with DAVID, BioMyn, AgBase, PrediSi, and Phobius. Compared to DAVID or BioMyn, identifications and annotations provided by ProteINSIDE were similar from monogastric proteins but more numerous and relevant for ruminants proteins. ProteINSIDE, thanks to SignalP, listed less proteins potentially secreted with a signal peptide than PrediSi and Phobius, in agreement with the low false positive rate of SignalP. In addition ProteINSIDE is the only resource that predicts proteins secreted by cellular processes that do not involve a signal peptide. Lastly, we reported the usefulness of ProteINSIDE to bring new biological hypotheses of research from proteomics data: the biological meaning of the uptake of adiponectin by the foetal muscle and a role for autophagy during ontogenesis of adipose and muscle

  7. Bovine Pericardial Patch Augmentation of One Insufficient Aortic Valve Cusp with Twenty-three-year Positive Clinical Follow-up Independent of the Patch Degeneration

    PubMed Central

    Evora, Paulo Roberto Barbosa; Arcêncio, Lívia; Evora, Patrícia Martinez; Menardi, Antônio Carlos; Chahud, Fernando

    2017-01-01

    Scientific progress shall ultimately boost the current acceptance level for conservative aortic valve surgery. The present text aimed to report the 23-year long-term follow-up of one patient operated with bovine pericardium cusp extension. Growing confidence in the efficacy of the operation will allow a more expeditious indication for surgical treatment, as is already the case in mitral valve repair. This change of attitude will certainly make it possible for patients to be sent for operation in mild aortic valve regurgitation. The present report reinforces the concept and highlights the impression that the aortic valvoplasty, independent of the progressive bovine pericardium degeneration, may positively change the natural history of the aortic valve insufficiency.

  8. Pulmonary artery patch for an inadequate donor atrial cuff in the absence of donor pericardium in lung transplantation.

    PubMed

    Sugimoto, Seiichiro; Yamane, Masaomi; Miyoshi, Kentaroh; Kurosaki, Takeshi; Otani, Shinji; Miyoshi, Shinichiro; Oto, Takahiro

    2017-03-01

    In cadaveric lung transplantation (LTx), a donor lung with an inadequate donor left atrial cuff is considered a "surgically marginal donor lung". The donor pericardium is commonly applied to reconstruct the inadequate donor left atrial cuff; however, in some cases, the donor pericardium is inadvertently removed during the lung procurement. We devised an alternative technique for reconstruction to overcome the absence of pericardium in a donor lung with an inadequate atrial cuff, using a patch of the donor pulmonary artery (PA) in single lung transplantation. In a recent case of lung transplantation in which the donor pericardium had been removed, we harvested a segment of the right PA distal to the main PA of the donor and used a PA patch to repair the inadequate donor left atrial cuff. No vascular complications were encountered in the recipient, who remains in good health after the transplantation.

  9. Bovine prolactin elevates hTF expression directed by a tissue-specific goat β-casein promoter through prolactin receptor-mediated STAT5a activation.

    PubMed

    Jiang, Shizhong; Ren, Zhaorui; Xie, Fei; Yan, Jingbin; Huang, Shuzhen; Zeng, Yitao

    2012-11-01

    Prolactin promotes the expression of exogenous human transferrin gene in the milk of transgenic mice. To elucidate this, a recombinant plasmid of bovine prolactin plus human transferrin vector was co-transfected into cultured murine mammary gland epithelial cells. Prolactin-receptor antagonist and shRNA corresponding to prolactin-receptor mRNA were added into the cell culture mixture to investigate the relations between prolactin-receptor and human transferrin expression after bovine prolactin inducement. Levels of human transferrin in the supernatants were increased under the presentation of bovine prolactin (from 1,076 ± 115 to 1,886 ± 114 pg/ml). With the treatment of prolactin-receptor antagonist or shRNA, human transferrin in cells was declined (1,886 ± 113 vs. 1,233 ± 85 pg/ml or 1,114 ± 75 pg/ml, respectively). An inverse correlation was found between the dosage of prolactin-receptor antagonist and expression level of human transferrin. Real-time qRT-PCR analysis showed that the relative level of signal transducer and activator of transcription 5a (STAT5a) transcript in transfected cells correlated with expression levels of human transferrin in the supernatant of the same cells. Bovine prolactin thus improved the expression of human transferrin through such a possible mechanism that bovine prolactin activated STAT5a transcription expression via combined with prolactin-receptor and suggest a potential utility of the bovine prolactin for efficient expression of valuable pharmaceutical proteins in mammary glands of transgenic animals.

  10. Five-year results of guided tissue regeneration in combination with deproteinized bovine bone (Bio-Oss) in the treatment of intrabony periodontal defects: a case series report.

    PubMed

    Stavropoulos, Andreas; Karring, Thorkild

    2005-12-01

    The aim of this study was to report on the clinical and radiographic results 5 years following treatment of intrabony defects with guided tissue regeneration (GTR) in combination with deproteinized bovine bone (DBB) (Bio-Oss). Fifteen patients, with at least one intrabony periodontal defect with probing pocket depth (PPD)>or=7 mm and radiographic presence of an intrabony component (IC)>or=4 mm, were treated with a PLA/PGA bioabsorbable membrane. Prior to placement of the membrane, the defect was filled with DBB impregnated with gentamicin sulfate 2 mg/ml. Standardized intraoral radiographs were taken prior to treatment and at the control examinations after 1 and 5 years. At baseline, the average PPD was 9.2+/-1.1 mm, and the average probing attachment level (PAL) was 10.1+/-1.6 mm; the radiographic bone level (RBL) was 10.4+/-2.45 mm, and an IC of 6.2+/-2.3 mm was present. One year after membrane placement, treatment had resulted in a PAL gain of 3.8+/-1.8 mm, a residual PPD of 4.2+/-1.3 mm, an RBL gain of 4.7+/-2.0 mm, and a residual IC of 2.1+/-1.2 mm. At the 5-year examination, two patients did not show up, and two patients had lost the treated tooth. However, both teeth were endodontically treated, and progressive periodontal destruction might not necessarily have been the reason for extraction. At the 5-year control (11 patients), the PAL gain was 4.1+/-1.6 mm, and the residual PPD was 4.6+/-1.2 mm; an RBL gain of 4.9+/-2.7 mm and a residual IC of 1.8+/-0.8 mm were observed. Statistically significant clinical improvements had occurred between baseline and the 1- and 5-year controls, whereas there were no significant differences between the 1- and 5-year results. The results of GTR with bioabsorbable membranes in combination with Bio-Oss in the treatment of periodontal intrabony defects are basically stable on a long-term basis.

  11. Incorporation of experimentally-derived fiber orientation into a structural constitutive model for planar collagenous tissues.

    PubMed

    Sacks, Michael S

    2003-04-01

    Structural constitutive models integrate information on tissue composition and structure, avoiding ambiguities in material characterization. However, critical structural information (such as fiber orientation) must be modeled using assumed statistical distributions, with the distribution parameters estimated from fits to the mechanical test data. Thus, full realization of structural approaches continues to be limited without direct quantitative structural information for direct implementation or to validate model predictions. In the present study, fiber orientation information obtained using small angle light scattering (SALS) was directly incorporated into a structural constitutive model based on work by Lanir (J. Biomech., v. 16, pp. 1-12, 1983). Demonstration of the model was performed using existing biaxial mechanical and fiber orientation data for native bovine pericardium (Sacks and Chuong, ABME, v.26, pp. 892-902, 1998). The structural constitutive model accurately predicted the complete measured biaxial mechanical response. An important aspect of this approach is that only a single equibiaxial test to determine the effective fiber stress-strain response and the SALS-derived fiber orientation distribution were required to determine the complete planar biaxial mechanical response. Changes in collagen fiber crimp under equibiaxial strain suggest that, at the meso-scale, fiber deformations follow the global tissue strains. This result supports the assumption of affine strain to estimate the fiber strains. However, future evaluations will have to be performed for tissue subjected to a wider range of strain to more fully validate the current approach.

  12. Effect of sample geometry on the apparent biaxial mechanical behaviour of planar connective tissues.

    PubMed

    Waldman, Stephen D; Lee, J Michael

    2005-12-01

    Mechanical testing methodologies developed for engineering materials may result in artifactual material properties if applied to soft planar connective tissues. The use of uniaxial tissue samples with high aspect ratios or biaxial samples with slender cruciform arms could lead to preferential loading of only the discrete subset of extracellular fibres that fully extend between the grips. To test this hypothesis, cruciform biaxial connective tissue samples that display distinctly different material properties (bovine pericardium, fish skin), as well as model textile laminates with predefined fibrous orientations, were repeatedly tested with decreasing sample arm lengths. With mechanical properties determined at the sample centre, results demonstrated that the materials appeared to become stiffer and less extensible with less slender sample geometries, suggesting that fibre recruitment increases with decreasing sample arm length. Alterations in the observed shear behaviour and rigid body rotation were also noted. The only truly reliable method to determine material properties is through in vivo testing, but this is not always convenient and is typically experimentally demanding. For the in vitro determination of the biaxial material properties, appropriate sample geometry should be employed in which all of the fibres contribute to the mechanical response.

  13. Quantitative criteria for the diagnosis of the congenital absence of pericardium by cardiac magnetic resonance.

    PubMed

    Macaione, F; Barison, A; Pescetelli, I; Pali, F; Pizzino, F; Terrizzi, A; Di Lisi, D; Novo, G; Todiere, G; Assennato, P; Novo, S; Aquaro, G D

    2016-03-01

    Congenital absence of the left ventricular pericardium (LCAP) is a rare and poorly known cardiac malformation. Cardiac Magnetic Resonance (CMR) is generally used for the diagnosis of LCAP because of its high soft tissue contrast, multiplanarity and cine capability, but the diagnosis is usually made by only qualitative criteria. The aim of the present study was to establish quantitative criteria for the accurate diagnosis of LCAP on CMR. We enrolled nine consecutive patients affected by LCAP (mean age 26±8years, 7 males), 13 healthy controls, 13 patients with dilated cardiomyopathy (DCM), 12 patients with hypertrophic cardiomyopathy (HCM) and 13 patients with right ventricular overload (RVO). All patients underwent CMR. The whole-heart volume was measured in end-systole and end-diastole. Whole-heart volume change (WHVC), was the systo-diastolic change of volume, expressed percentage of the end-diastolic volume. The angle of clockwise-rotation of the heart was also measured in the end-diastolic phase of the axial cine stack. The WHVC was significantly higher in LCAP (21.9±5.4), compared to healthy subjects (8.6±2.4, p<0.001), DCM (7.1±1.8, p<0.001), HCM (9.3±2.4, p<0.001) and RVO (8±2.4, p<0.001). The clockwise-rotation was significantly higher in LCAP (76±14°) than healthy controls (40±11°, p<0.001), DCM (41±5°, p<0.001), HCM (30±6°, p<0.001) and RVO (49±8°, p<0.001). WHVC had the highest sensitivity (100%) and specificity (100%) for diagnosing LCAP, using a threshold of >13%. In LCAP the systo-diastolic WHVC was significantly higher than controls, DCM, HCM and RVO patients and resulted an optimal quantitative criteria for the diagnosis of LCAP. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  14. Comparison of stability of viral nucleic acid in different tissues and under different conditions in samples collected from fetuses infected with bovine viral diarrhea virus.

    USDA-ARS?s Scientific Manuscript database

    Accurate diagnosis of bovine viral diarrhea virus (BVDV) induced reproductive disease is important to herd health management and BVDV control programs. Diagnosing BVDV, by polymerase chain reaction (PCR), as a cause of reproductive disease may be problematic because viral nucleic acid may be degrade...

  15. Involvement of Connective Tissue Growth Factor (CTGF) in Insulin-like Growth Factor-I (IGF1) Stimulation of Proliferation of a Bovine Mammary Epithelial Cell Line

    USDA-ARS?s Scientific Manuscript database

    Insulin-like growth factor I (IGF1) plays an important role in mammary gland development and lactation in part by stimulating proliferation of the milk-producing epithelial cells. In this study, we used the bovine mammary epithelial cell line MAC-T cells as a model to understand the mechanism by whi...

  16. Computed Tomography-guided Pericardiocentesis: An alternative approach for accessing the pericardium.

    PubMed

    Melvan, John Nicholas; Madden, David; Vasquez, Julio C; DeLaRosa, Jacob

    2016-07-01

    Pericardial effusions compress the heart, decrease cardiac output, and lead to haemodynamic collapse. Ultrasound (US)-guided pericardiocentesis is the gold standard for treating pericardial effusions. Recently, the incorporation of computed tomography (CT) guidance has increased patient safety while entering the pericardium. Despite the superior performance of CT-guided pericardiocentesis in smaller, complex effusions, this procedure is not routinely performed by cardiologists and surgeons. Unlike those with an intact pericardium, patients with mediastinal trauma, pericardial adhesions, temporary pacing wires, and vascular conduits are high risk for pericardiocentesis. Tamponade physiology also increases patient susceptibility to the hypotensive effects of anaesthesia during surgical drainage. Here we illustrate the technique of CT-guided pericardiocentesis and demonstrate its application in specific clinical scenarios. We conclude that CT-guided pericardiocentesis provides a useful, alternative strategy for treating cardiac tamponade in high risk patients.

  17. The 2015 WHO Classification of Tumors of the Heart and Pericardium.

    PubMed

    Burke, Allen; Tavora, Fabio

    2016-04-01

    This article reviews the nomenclature of benign and malignant neoplasm of the heart and pericardium in the 4th edition of the World Health Organization's Classification, with emphasis on differences since the 3rd edition of 2004. The tumours are divided into benign, malignant, and intermediate tumors of uncertain behavior, with separate sections on germ cell tumours and tumors of the pericardium. There are important updates in the sarcoma classification, with emphasis on the most common site, the left atrium. The importance of the new genetic finding in cardiac myxomas, namely somatic mutations in the PRKAR1A gene underscores the importance of this alteration in the pathogenesis of these tumors. Challenges on the classification of each entity are discussed. Published by Elsevier Inc.

  18. Teflon-buttressed sutures plus pericardium patch repair left ventricular rupture caused by radiofrequency catheter ablation

    PubMed Central

    Cao, Hao; Zhang, Qi; He, Yanzhong; Feng, Xiaodong; Liu, Zhongmin

    2016-01-01

    Abstract Background: Cardiac rupture often occurs after myocardial infarction or chest trauma with a high mortality rate. However, left ventricular rupture caused by radiofrequency catheter ablation (RFCA) is extremely rare. Methods: We describe a case of a 61-year-old male who survived from left ventricular rupture caused by a RFCA procedure for frequent ventricular premature contractions. Surgical exploration with cardiopulmonary bypass (CPB) was performed when the signs of cardiac tamponade developed 7 hours after the ablation surgery. Results: Teflon-buttressed sutures of the tear in the left ventricular posterolateral wall and pericardium patch applied to the contusion region on the wall repaired the rupture safely and effectively. Conclusion: Timely surgical intervention under CPB facilitated the survival of the patient. Teflon-buttressed sutures plus pericardium patch achieved the successful repair of the rupture. PMID:27661047

  19. Congenital complete absence of pericardium in a young woman with non-specific symptoms.

    PubMed

    Bueno Palomino, Antonio; Palomar Estrada, Alberto; Crespín Crespín, Manuel; García Fuertes, Daniel

    2014-04-01

    Congenital absence of the pericardium is a very rare entity that is usually asymptomatic and hence difficult to diagnose. However, cases of sudden death have been reported in patients with partial pericardial defects (even asymptomatic ones), and such patients require surgical treatment. We report the case of a 17-year-old patient with complete pericardial agenesis (diagnosed by chance during a cardiological consultation) and briefly review the radiological findings of this entity.

  20. [Atrioventricular defect with 2 valves associated with absence of the pericardium. Report of a case].

    PubMed

    Miranda-Chávez, Irma; Muñoz-Castellanos, Luis; Kimura-Hayama, Erick; Vidal-Rojo, Paola; Figueroa-Solano, Javier; de Micheli Serra, Alfredo; Buendía-Hernández, Alfonso

    2013-01-01

    We present a case of a patient with a cardiac malformation that represents a form of embryo-anatomical transition of an atrioventricular septal defect between a 2 valves form to a common valve form. This entity was associated with pericardium absence. Throughout several studies we have precisely established a diagnostic sequence by determining the adequate contribution of each method and we have been able to clear out the proper nomenclature of the atrioventricular cushion defect.

  1. [Congenital Defect of the Left Pericardium with Spontaneous Pneumothorax;Report of a Case].

    PubMed

    Murasawa, Masaki; Yoshizawa, Masatoshi; Ishida, Hisao; Kuwabara, Masayoshi

    2016-08-01

    Congenital cardiac membrane deficiency is a relatively rare condition. Here, we report a case of congenital cardiac membrane deficiency that manifested as left spontaneous pneumothorax. A 72-year-old man was hospitalized for recurrence of the spontaneous pneumothorax. Computed tomography findings led us to suspect pericardial deficits, and the perioperative findings during thoracoscopic surgery for the pneumothorax confirmed complete absence of the left pericardium. We resected a lung cyst but did not treat the pericardial deficit. The patient's postoperative course was uneventful.

  2. Congenital diaphragmatic hernia with concurrent aplasia of the pericardium in a foal.

    PubMed

    Tăbăran, Alexandru-Flaviu; Nagy, Andras Laszlo; Cătoi, Cornel; Morar, Iancu; Tăbăran, Alexandra; Mihaiu, Marian; Bolfa, Pompei

    2015-12-30

    In veterinary medicine congenital abnormalities of the diaphragm and pericardium are rare, idiopathic malformations, being reported mainly in dogs. This report documents an unusual case of developmental defects in a foal consisting of diaphragmatic hernia concurrent with pericardial aplasia. Following a normal delivery, a full term, female Friesian stillborn foal with the placenta was presented for necropsy. External morphological examination indicated a normally developed foal. At necropsy, a large oval defect (approximately 20 × 15 cm in size) was observed in the left-dorsal side of the diaphragm (left lumbocostal triangle). This defect allowed the intestinal loops, spleen and partially the liver to translocate into the thorax. The loops of the left ascending colon, including the pelvic flexure and partially the small intestine covered the cranial and dorsal posterior parts of the heart due to the complete absence of the left pericardium. The remaining pericardium presented as a white, semi-transparent strip, partially covering the right side of the heart. The left lung and the main bronchus were severely hypoplastic to approximately one-fifth the size of their right homologue. The intermediate part of the liver, containing mainly the enlarged quadrate lobe was translocated in the thorax, severely enlarged and showed marked fibrosis. Histologically in the herniated lobes we diagnosed hepatic chronic passive congestion, telangiectasia and medial hypertrophy of blood vessels. Concomitant malformation involving diaphragmatic hernia and pericardial aplasia in horses have not been previously reported. Moreover, this is the first case describing pericardial aplasia in horse.

  3. Transport properties of the mesothelium and interstitium measured in rabbit pericardium.

    PubMed

    Tang, Sonja M Moe; Lai-Fook, Stephen J

    2005-11-01

    The contribution of the pleural mesothelium to pleural liquid and protein transport is still vigorously debated. Recent in vitro studies of stripped pleural membrane and free-standing pericardium have demonstrated active ion solute coupled transport of liquid and transcytosis of protein. However, the relative contribution of the passive transport properties of the pleural mesothelium compared to the pleural interstitium has not been extensively studied. In in vitro studies, we measured the albumin diffusion coefficient, reflection coefficient, hydraulic conductivity and electrical resistance of rabbit pericardium. We used two techniques, treatment with 40 muM nocodazole and a 1-min hypotonic cell lysis with distilled water, to eliminate the effect of the two mesothelial layers on diffusional and hydraulic resistances. Each technique increased the albumin diffusion coefficient and hydraulic conductivity 3- to 4-fold. In hydraulic conductivity experiments using tracer 125I-albumin, nocodazole reduced the reflection coefficient to zero, rendering the pericardium completely permeable to albumin. We applied the cell-lysis technique to the pleural and pericardial mesothelium in sequence to evaluate the separate contribution of each mesothelium. Both diffusional and hydraulic resistances, but not electrical resistance, of the mesothelium were overestimated by the cell-lysis technique. The pleural mesothelium contributed at most 30% of diffusional resistance, 10% of hydraulic resistance and 14% of electrical resistance of the total pericardial resistances. We conclude that the pleural mesothelium is not the primary barrier to protein diffusion or bulk flow of liquid from the pericardial microcirculation to the pleural liquid.

  4. A 2D FE model of the heart demonstrates the role of the pericardium in ventricular deformation.

    PubMed

    Gibbons Kroeker, Carol A; Adeeb, Samer; Tyberg, John V; Shrive, Nigel G

    2006-11-01

    During pulmonary artery constriction (PAC), an experimental model of acute right ventricular (RV) pressure overload, the interventricular septum flattens and inverts. Finite element (FE) analysis has shown that the septum is subject to axial compression and bending when so deformed. This study examines the effects of acute PAC on the left ventricular (LV) free wall and the role the pericardium may play in these effects. In eight open-chest anesthetized dogs, LV, RV, aortic, and pericardial pressures were recorded under control conditions and with PAC. Model dimensions were derived from two-dimensional echocardiography minor-axis images of the heart. At control (pericardium closed), FE analysis showed that the septum was concave to the LV; stresses in the LV, RV, and septum were low; and the pericardium was subject to circumferential tension. With PAC, RV end-diastolic pressure exceeded LV pressure and the septum inverted. Compressive stresses developed circumferentially in the septum out to the RV insertion points, forming an arch-like pattern. Sharp bending occurred near the insertion points, accompanied by flattening of the LV free wall. With the pericardium open, the deformations and stresses were different. The RV became much larger, especially with PAC. With PAC, the arch-like circumferential stresses still developed in the septum, but their magnitudes were reduced, compared with the pericardium-closed case. There was no free wall inversion and flattening was less. From these FE results, the pericardium has a significant influence on the structural behavior of the septum and the LV and RV free walls. Furthermore, the deformation of the heart is dependent on whether the pericardium is open or closed.

  5. Morphological characteristics of cultured fresh and thawed pericardium cells.

    PubMed

    Maslova, Olga; Fedevych, Oleg; Shuvalova, Nadiia; Deryabina, Olena; Zhovnir, Volodymyr; Novak, Miroslav; Kruzliak, Peter

    2016-06-01

    The need for selection of the optimal material for the manufacturing of cardio-patches can be resolved by the use of cryostored autologous pericardial tissue. This short communication is a concise fragment of a large-scale research and demonstrates only the efficiency of cell culturing before and after pericardial preservation in the low temperature conditions.

  6. Biological and mechanical evaluation of a Bio-Hybrid scaffold for autologous valve tissue engineering.

    PubMed

    Jahnavi, S; Saravanan, U; Arthi, N; Bhuvaneshwar, G S; Kumary, T V; Rajan, S; Verma, R S

    2017-04-01

    Major challenge in heart valve tissue engineering for paediatric patients is the development of an autologous valve with regenerative capacity. Hybrid tissue engineering approach is recently gaining popularity to design scaffolds with desired biological and mechanical properties that can remodel post implantation. In this study, we fabricated aligned nanofibrous Bio-Hybrid scaffold made of decellularized bovine pericardium: polycaprolactone-chitosan with optimized polymer thickness to yield the desired biological and mechanical properties. CD44(+), αSMA(+), Vimentin(+) and CD105(-) human valve interstitial cells were isolated and seeded on these Bio-Hybrid scaffolds. Subsequent biological evaluation revealed interstitial cell proliferation with dense extra cellular matrix deposition that indicated the viability for growth and proliferation of seeded cells on the scaffolds. Uniaxial mechanical tests along axial direction showed that the Bio-Hybrid scaffolds has at least 20 times the strength of the native valves and its stiffness is nearly 3 times more than that of native valves. Biaxial and uniaxial mechanical studies on valve interstitial cells cultured Bio-Hybrid scaffolds revealed that the response along the axial and circumferential direction was different, similar to native valves. Overall, our findings suggest that Bio-Hybrid scaffold is a promising material for future development of regenerative heart valve constructs in children. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Nanosecond laser ablation enhances cellular infiltration in a hybrid tissue scaffold.

    PubMed

    Jahnavi, S; Arthi, N; Pallavi, S; Selvaraju, C; Bhuvaneshwar, G S; Kumary, T V; Verma, R S

    2017-08-01

    Hybrid tissue engineered (HTE) scaffolds constituting polymeric nanofibers and biological tissues have attractive bio-mechanical properties. However, they suffer from small pore size due to dense overlapping nanofibers resulting in poor cellular infiltration. In this study, using nanosecond (ns) laser, we fabricated micro-scale features on Polycaprolactone (PCL)-Chitosan (CH) nanofiber layered bovine pericardium based Bio-Hybrid scaffold to achieve enhanced cellular adhesion and infiltration. The laser energy parameters such as fluence of 25J/cm(2), 0.1mm instep and 15 mark time were optimized to get structured microchannels on the Bio-Hybrid scaffolds. Laser irradiation time of 40μs along with these parameters resulted in microchannel width of ~50μm and spacing of ~35μm between adjacent lines. The biochemical, thermal, hydrophilic and uniaxial mechanical properties of the Bio-Hybrid scaffolds remained comparable after laser ablation reflecting extracellular matrix (ECM) stability. Human umbilical cord mesenchymal stem cells and mouse cardiac fibroblasts seeded on these laser-ablated Bio-Hybrid scaffolds exhibited biocompatibility and increased cellular adhesion in microchannels when compared to non-ablated Bio-Hybrid scaffolds. These findings suggest the feasibility to selectively ablate polymer layer in the HTE scaffolds without affecting their bio-mechanical properties and also describe a new approach to enhance cellular infiltration in the HTE scaffolds. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Induction of peroxisomal proliferator-activated receptor gamma and peroxisomal proliferator-activated receptor gamma coactivator 1 by unsaturated fatty acids, retinoic acid, and carotenoids in preadipocytes obtained from bovine white adipose tissue1,2.

    PubMed

    García-Rojas, P; Antaramian, A; González-Dávalos, L; Villarroya, F; Shimada, A; Varela-Echavarría, A; Mora, O

    2010-05-01

    The importance of dietary fat components, such as fatty acids, in the expression of multiple genes is clear. In the case of beef cattle, fat in the form of fatty acids (saturated or unsaturated), vitamin A (mainly retinoic acid), or carotenoids (beta-carotene and lutein) is obtained from dietary feed or pasture. The aim of this work was to study the effect of fatty acids (phytanic and pristanic acids), vitamin A (all-trans and 9-cis retinoic acid), and carotenoids (beta-carotene and lutein) on the expression of PPARgamma and its coactivator PGC-1alpha during differentiation of bovine white adipose tissue. Samples were collected at slaughter from subcutaneous adipose tissue and processed in a solution containing type II collagenase for 2 h at 37 degrees C. Cells were resuspended in basal medium, Dulbecco's modified Eagle's medium containing 5% fetal bovine serum, plated on 24-well culture plates at a density of 1 x 10(4) cells/cm(2), and incubated at 37 degrees C in a 5% CO(2) atmosphere. Preadipocyte differentiation after reaching confluence was induced by various treatments: rosiglitazone (20 microM); unsaturated fatty acids: phytanic acid (25, 50, 100 microM) and pristanic acid (25, 50, 100 microM); retinoids: 9-cis retinoic acid (0.5, 0.75, 1 microM) and all-trans retinoic acid (0.5, 0.75, 1 microM); and carotenoids: beta-carotene (10, 20, 30 microM) and lutein (10, 20, 30 microM). Expression of PPARgamma and PGC-1alpha was measured in differentiated cells. Phytanic acid, all-trans retinoic acid, and 9-cis retinoic acid were the best activators of PPARgamma expression, and the combination of 9-cis and all-trans retinoic acid was the best activator of PGC-1alpha expression (P < 0.05). Therefore, these are powerful agents for the promotion of bovine adipogenesis and constitute promising compounds to be used in bovine fattening.

  9. Acute infection with bovine viral diarrhea virus of low or high virulence leads to depletion and redistribution of WC1(+) γδ T cells in lymphoid tissues of beef calves.

    PubMed

    Palomares, Roberto A; Sakamoto, Kaori; Walz, Heather L; Brock, Kenny V; Hurley, David J

    2015-10-15

    The objective of this study was to determine the abundance and distribution of γδ T lymphocytes in lymphoid tissue during acute infection with high (HV) or low virulence (LV) non-cytopathic bovine viral diarrhea virus (BVDV) in beef calves. This study was performed using tissue samples from a previous experiment in which thirty beef calves were randomly assigned to 1 of 3 groups: LV [n=10; animals inoculated intranasally (IN) with LV BVDV-1a (strain SD-1)], HV [n=10; animals inoculated IN with HV BVDV-2 (strain 1373)], and control (n=10; animals inoculated with cell culture medium). On day 5 post inoculation, animals were euthanized, and samples from spleen and mesenteric lymph nodes (MLN) were collected to assess the abundance of WC1(+) γδ T cells. A higher proportion of calves challenged with BVDV showed signs of apoptosis and cytophagy in MLN and spleen samples compared to the control group. A significantly lower number of γδ T cells was observed in spleen and MLN from calves in HV and LV groups than in the control calves (P<0.05). In conclusion, acute infection with HV or LV BVDV resulted in depletion of WC1(+) γδ T cells in mucosal and systemic lymphoid tissues at five days after challenge in beef calves. This reduction in γδ T cells in the studied lymphoid tissues could be also due to lymphocyte trafficking to other tissues.

  10. Angiosarcoma of pericardium: a report of two cases.

    PubMed

    Bhattacharya, P; Singh, Paramjit; Bahal, A; Gulati, D; Tevatia, M S; Mehta, A; Muttagikar, M P

    2007-10-01

    Angiosarcomas are uncommon malignant neoplasms characterized by rapidly proliferating extensively infiltrating anaplastic cells derived from blood vessels and lining irregular, blood-filled spaces. The cells manifest many of the functional and morphological properties of normal endothelium. They are collectively one of the rarest forms of soft tissue neoplasms. Here we present two cases of pericardial angiosarcoma, one of them showing widespread dissemination, which caused considerable diagnostic dilemma and the diagnosis could only be established very late in their course of disease.

  11. Characterisation of human skin impedance at acupuncture point PC4 Ximen and pericardium meridian using the four-electrode method.

    PubMed

    Rezaei, Shima; Khorsand, Ali; Jamali, Jamshid

    2012-06-01

    Traditional Chinese medicine offers several theories to explain the mechanism of acupuncture. One of these theories proposes that acupuncture points and meridians have unique electrical properties and their electrical skin impedance is lower than surrounding areas. The aim of this study was to evaluate the differences in electrical skin impedance between PC4 and the pericardium meridian compared with the surrounding areas. Eighteen healthy subjects (10 women) were recruited to participate in the study. An impedance meter based on the four-electrode technique was designed specifically for the study. Twenty-five points were marked on the skin: one on the point PC4, four others on the pericardium meridian and 20 points around it. The electrical impedance of each point was measured with the four-electrode device. The mean electrical skin impedance at PC4 was significantly different from the 20 of the surrounding points but not significantly different from the four adjacent points. The mean skin impedance of the five points over the pericardium meridian was significantly different from that of parallel rows of points using repeated measures analysis of variance (p<0.001) Within the possible limits of this measurement technique, skin impedance along the pericardium meridian is lower than surrounding areas, supporting the idea of different properties of the pericardium meridian compared with the control areas. Evidence on skin impedance at PC4 is inconclusive and further studies are needed.

  12. A deep learning approach to estimate chemically-treated collagenous tissue nonlinear anisotropic stress-strain responses from microscopy images.

    PubMed

    Liang, Liang; Liu, Minliang; Sun, Wei

    2017-09-20

    Biological collagenous tissues comprised of networks of collagen fibers are suitable for a broad spectrum of medical applications owing to their attractive mechanical properties. In this study, we developed a noninvasive approach to estimate collagenous tissue elastic properties directly from microscopy images using Machine Learning (ML) techniques. Glutaraldehyde-treated bovine pericardium (GLBP) tissue, widely used in the fabrication of bioprosthetic heart valves and vascular patches, was chosen to develop a representative application. A Deep Learning model was designed and trained to process second harmonic generation (SHG) images of collagen networks in GLBP tissue samples, and directly predict the tissue elastic mechanical properties. The trained model is capable of identifying the overall tissue stiffness with a classification accuracy of 84%, and predicting the nonlinear anisotropic stress-strain curves with average regression errors of 0.021 and 0.031. Thus, this study demonstrates the feasibility and great potential of using the Deep Learning approach for fast and noninvasive assessment of collagenous tissue elastic properties from microstructural images. In this study, we developed, to our best knowledge, the first Deep Learning-based approach to estimate the elastic properties of collagenous tissues directly from noninvasive second harmonic generation images. The success of this study holds promise for the use of Machine Learning techniques to noninvasively and efficiently estimate the mechanical properties of many structure-based biological materials, and it also enables many potential applications such as serving as a quality control tool to select tissue for the manufacturing of medical devices (e.g. bioprosthetic heart valves). Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  13. Quantification and determination of spread mechanisms of bovine viral diarrhoea virus in blood and tissues from colostrum-deprived calves during an experimental acute infection induced by a non-cytopathic genotype 1 strain.

    PubMed

    Pedrera, M; Gómez-Villamandos, J C; Molina, V; Risalde, M A; Rodríguez-Sánchez, B; Sánchez-Cordón, P J

    2012-10-01

    To detect and monitor the sequential changes in virus levels, a reverse transcription quantitative real-time polymerase chain reaction assay using a TaqMan probe was carried out on frozen blood and tissues samples collected from calves experimentally infected with a non-cytopathic Bovine viral diarrhoea virus (BVDV) genotype 1 strain. Blood samples were collected among days 1-14 post-inoculation (p.i). On day 3 p.i, viral RNA was detected in blood samples from six of the eight inoculated animals. Viral RNA was detected in all remaining inoculated animals between 5 and 12 days p.i. The levels of viral RNA increased along the experiment, with a maximal peak between 6 and 9 days p.i. Analysis of virus load in tissues collected from calves euthanized on days 3, 6, 9 and 14 p.i displayed that BVDV was detected on day 3 p.i, being especially abundant in tonsils and ileocaecal valve, highlighting the role of tonsils as the main earliest viral replication sites as well as the principal source for virus spread to other lymphoid tissues and visceral organs. Coinciding with the highest viraemia levels, the highest viral loads were recorded at 9 days p.i. in tonsils, ileal lymph nodes, distal ileum and spleen, showing the main role of these secondary lymphoid organs in the pathogenic mechanisms of BVDV. However, virus levels in the liver and lung increased only towards the end of the infection. This fact could influence in the appearance of bovine respiratory diseases because of the capacity of BVDV for enhancing susceptibility to secondary infections.

  14. Indications and immediate and long-term results of a novel pericardium covered stent graft: Consecutive 5 year single center experience.

    PubMed

    Secco, Gioel Gabrio; Serdoz, Roberta; Kilic, Ismail Dogu; Caiazzo, Gianluca; Mattesini, Alessio; Parisi, Rosario; De Luca, Giuseppe; Pistis, Gianfranco; Marino, Paolo Nicola; Di Mario, Carlo

    2016-03-01

    The use of covered stent grafts during percutaneous coronary intervention (PCI) is a life saving solution to seal acute iatrogenic vessel rupture. However, the presence of an impenetrable mechanical barrier is also appealing during treatment of friable coronary plaques but the synthetic PTFE-membrane that might trigger excessive neointimal proliferation has limited its elective-use. Pericardium tissue may offer an appealing "natural" alternative. Aim of our study is to report the consecutive 5-year single center experience with the use of pericardium-covered stents (PCS) (ITGI-Medical, Israel) in a variety of emergency and elective applications. Nineteen consecutive patients undergoing implantation of PCS at the Royal Brompton in the last 5-years. Reasons for PCS implantation included treatment of degenerated vein grafts, large coronary aneurysms, and acute iatrogenic vessel rupture. Angiographic success, defined as the ability of the device to be deployed in the indexed lesion with no contrast extravasation with residual angiographic stenosis <30% and a final thrombolysis in myocardial infarction (TIMI)-3 flow was achieved in all cases. Procedural success, defined as the achievement of angiographic success without any major adverse cardiovascular event (MACE) was achieved in 94.7% of patients. In-stent restenosis (ISR) was observed in 26.3% and all patients underwent successful target vessel revascularization with DES (mean time to restenosis 9.0 ± 4.0 months). At a mean follow-up of 32.5 ± 23.3 months no acute or late stent thrombosis was observed. PCSs were effective in the treatment of friable embolization-prone coronary plaques, sealing of acute iatrogenic vessel rupture and exclusion of large aneurysms with no thrombosis but high target lesion revascularization. © 2015 Wiley Periodicals, Inc.

  15. Hyperenhancement of the Pericardium on Cardiac Magnetic Resonance Imaging: A Marker of Acute Inflammation and Neovascularization or a Chronic Fibrotic State.

    PubMed

    Mullen, Liam; Chew, Pei Gee; Frost, Frederick; Ahmed, Ayesha; Khand, Aleem

    2016-01-01

    In cardiac magnetic resonance imaging, hyperenhancement of the pericardium post gadolinium administration in acute chest pain often signifies pericarditis with an acute inflammatory response and neovascularization. In the context of constrictive pericarditis, case series have indicated that the intensity of hyperenhancement and the thickness of the pericardium imply reversibility of the physiology of the constrictive pericarditis. We present a case of intense hyperenhancement and marked thickening of the pericardium in a patient with constrictive pericarditis with antecedent chest pain. Surgical resection of the pericardium and microscopy revealed a chronic fibrotic state with no evidence of inflammation or neovascularization, thus clarifying the failure of initial medical/anti-inflammatory treatment. Our case highlights the fact that hyperenhancement of the pericardium post gadolinium is non-specific for histology and does not necessarily imply the reversibility of pericardial constriction. © 2016 S. Karger AG, Basel.

  16. Stabilized Collagen Scaffolds for Heart Valve Tissue Engineering

    PubMed Central

    Tedder, Mary E.; Liao, Jun; Weed, Benjamin; Stabler, Christopher; Zhang, Henry; Simionescu, Agneta

    2009-01-01

    Scaffolds for heart valve tissue engineering must function immediately after implantation but also need to tolerate cell infiltration and gradual remodeling. We hypothesized that moderately cross-linked collagen scaffolds would fulfill these requirements. To test our hypothesis, scaffolds prepared from decellularized porcine pericardium were treated with penta-galloyl glucose (PGG), a collagen-binding polyphenol, and tested for biodegradation, biaxial mechanical properties, and in vivo biocompatibility. For controls, we used un-cross-linked scaffolds and glutaraldehyde-treated scaffolds. Results confirmed complete pericardium decellularization and the ability of scaffolds to encourage fibroblast chemotaxis and to aid in creation of anatomically correct valve-shaped constructs. Glutaraldehyde cross-linking fully stabilized collagen but did not allow for tissue remodeling and calcified when implanted subdermally in rats. PGG-treated collagen was initially resistant to collagenase and then degraded gradually, indicating partial stabilization. Moreover, PGG-treated pericardium exhibited excellent biaxial mechanical properties, did not calcify in vivo, and supported infiltration by host fibroblasts and subsequent matrix remodeling. In conclusion, PGG-treated acellular pericardium is a promising scaffold for heart valve tissue engineering. PMID:18928400

  17. A simple method of aortic valve reconstruction with fixed pericardium in children

    PubMed Central

    Hosseinpour, Amir-Reza; González-Calle, Antonio; Adsuar-Gómez, Alejandro; Santos-deSoto, José

    2013-01-01

    Aortic valve reconstruction with fixed pericardium may occasionally be very useful when treating children with aortic valve disease. This is because diseased aortic valves in children are sometimes too dysmorphic for simple repair without the addition of material, their annulus may be too small for a prosthesis, and the Ross operation may be precluded due to other congenital anomalies such as pulmonary valvar or coronary malformations. Such reconstruction is usually technically demanding and requires much precision. We describe a simple alternative method, which we have carried out in 3 patients, aged 1 week, 3 years and 12 years, respectively, with good early results. PMID:23343835

  18. Multiresidue method for identification and quantification of avermectins, benzimidazoles and nitroimidazoles residues in bovine muscle tissue by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) using a QuEChERS approach.

    PubMed

    Silva, Guilherme Resende da; Lima, Josefa Abucater; Souza, Leonardo Francisco de; Santos, Flávio Alves; Lana, Mary Ane Gonçalves; Assis, Débora Cristina Sampaio de; Cançado, Silvana de Vasconcelos

    2017-08-15

    A quantitative and confirmatory multiresidue method for determining the presence of avermectins, benzimidazoles and nitroimidazoles in bovine muscle tissue by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was developed, optimized and validated, using a QuEChERS extraction. The evaluated performance parameters were linearity, selectivity, matrix effect, decision limits (CCα), detection capability (CCβ), limits of detection (LOD), limits of quantification (LOQ), accuracy, precision and robustness. The validated method exhibited linearity with coefficient of determination (R(2)) higher than 0.90 in the working range from 0.5 to 2.0 times the maximum residue limit (MRL) or the minimum required performance level (MRPL) for the studied analytes, except for closantel, for which the linear study range was defined from 50 to 200µgkg(-1). The method was selective in the presence of macrolides and lincosamides for all the studied analytes. The LOD varied from 0.007 to 66.715µgkg(-1), whereas LOQ values ranging from 0.011 to 113.674µgkg(-1) were found. The results of the evaluation of the accuracy and precision were satisfactory for all the studied analytes, and according to the assessment of the robustness, the method was not robust only for the analytes abamectin, moxidectin, doramectin fenbendazole sulfone, closantel, thiabendazole, hydroxyl-metronidazole and ronidazole. The performance parameters demonstrated total method adequacy for the detection and quantification of avermectins, benzimidazoles and nitroimidazoles residues in bovine muscle tissues. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Endovascular Treatment of Internal Carotid and Vertebral Artery Aneurysms Using a Novel Pericardium Covered Stent

    PubMed Central

    Vulev, I.; Klepanec, A.; Bazik, R.; Balazs, T.; Illes, R.; Steno, J.

    2012-01-01

    Summary Intracranial aneurysm is a fairly common (often asymptomatic) condition. Subarachnoid hemorrhage associated with aneurysmal rupture is a potentially lethal event with a mortality rate as high as 50 percent and a high rate of disability among those who survive the initial hemorrhage, such that recently published guidelines support treatment of intracerebral aneurysms. The current treatment options include surgical clipping and endovascular treatment, but these are not without significant problems. Despite the trend toward endovascular treatment the rate of recurrence and complications is high. Current published evidence of the use of covered stent is limited to stents covered with polytetrafluoroethylene. It is now recognized that mammalian extracellular matrix represents an excellent scaffold material suitable for many therapeutic applications and glutaraldehyde treated pericardium has been widely used for many years due to its desirable features such as low immunogenicity and durability. This report describes the first published experience with the Aneugraft Pericardium Covered Stent (ITGI Medical, OR Akiva, Israel) in the treatment of internal carotid and vertebral artery aneurysms in three patients. In all three cases, the implantation of this novel device has resulted in successful closure of aneurysms. PMID:22681731

  20. Congenital partial absence of the pericardium presenting with a rare concurrent abnormality of vascular ring diagnosed by cardiac magnetic resonance imaging

    PubMed Central

    Sani, Zahra Alizadeh; Savand-Roomi, Zahra; Vojdanparast, Mohammad; Sarafan, Shadi; Seifi, Azin; Nezafati, Pouya

    2016-01-01

    Congenital absence of the pericardium is a rare abnormality that can be diagnosed by cardiac imaging procedures. A 49-year-old male needed medical attention due to the appearance of palpitation with a systolic murmur, and a notable aortic arch deviation was seen in the chest X-ray. In the echocardiogram, a poor echo window was detected. A cardiac magnetic resonance imaging (MRI) showed a rare concomitant anomaly of partial absence of the pericardium including a rare defect of the right-sided aortic arch. Using cardiac MRI, the pericardium can be easily visualized, and thus, its absence more easily detected, aiding appropriate clinical decision-making. PMID:28217641

  1. A novel in situ polymerase chain reaction hybridisation assay for the direct detection of bovine herpesvirus type 5 in formalin-fixed, paraffin-embedded tissues.

    PubMed

    Cardoso, Tereza C; Gomes, Deriane E; Ferrari, Heitor F; Silva-Frade, Camila; Rosa, Ana C G; Andrade, Alexandre L; Luvizotto, Maria Cecília R

    2010-02-01

    An in situ polymerase chain reaction (IS-PCR) hybridisation assay was carried out on the brains of 20 cattle infected naturally with bovine herpesvirus type 5 (BoHV-5). Sections from the olfactory bulb and the frontal cortex of each sample were analysed using IS-PCR followed by hybridisation targeting the BoHV-5 US9 gene using a biotinylated primer. Each of the IS-PCR and hybridisation steps was optimised, and three different methods for detecting the virus were used. No false positive signals were observed in any negative control sample (n=20), resulting in a specificity of 100%. The results of IS-PCR hybridisation analysis of the olfactory bulb and the frontal cortex be compared directly with the results obtained using virus isolation, and the specificity and sensitivity were calculated. The most suitable method of visualisation was the peroxidase/3'-3-diaminobenzidine (DAB) detection system coupled with the use of the fluorescent dye Cy3. Using either of these methods, 80% of the positive samples (16 out of 20 samples) were identified using olfactory bulb sections. This is the first report using IS-PCR hybridisation for the direct detection of BoHV-5 DNA in clinical samples, and it provides an additional method for veterinary virology.

  2. Singlet Fission Reaction of Light-Exposed β-Carotene Bound to Bovine Serum Albumin. A Novel Mechanism in Protection of Light-Exposed Tissue by Dietary Carotenoids.

    PubMed

    Chang, Hui-Ting; Chang, Yu-Qiang; Han, Rui-Min; Wang, Peng; Zhang, Jian-Ping; Skibsted, Leif H

    2017-07-26

    We have attempted to investigate the role of carotenoids (Car) in protecting pigment-protein complexes against light-induced degradation. Upon direct photoexcitation of β-carotene (β-Car), nanosecond flash photolysis and femtosecond time-resolved spectroscopy detected a substantial population of triplet states for β-Car aggregates associated with bovine serum albumin (BSA) or dispersed in aqueous phase with 10% tetrahydrofuran (THF), but none were observed for monomeric β-Car in neat THF. The direct photogeneration of triplet states was on the time scale of <1 ps, indicating that the underlying reaction mechanism was singlet fission (SF). Efficient triplet-triplet annihilation in the time regime from picoseconds to microseconds resulted in a <1 μs triplet lifetime for β-Car aggregates, in contrast to a 20 μs lifetime for monomeric β-Car as determined by anthracene-sensitized flash photolysis. The short-lived triplet excitations of β-Car aggregates associated with BSA or dispersed in aqueous phase were found to be insensitive to the presence of oxygen, which are considered to be important for the protection of both protein and carotenoid against light-induced degradation via reaction with oxidative species.

  3. Development of a HS-SPME-GC-MS/MS Method for the Quantitation of Thymol and Carvacrol in Bovine Matrices and To Determine Residue Depletion in Milk and Tissues.

    PubMed

    Armorini, Sara; Yeatts, James E; Mullen, Keena A E; Mason, Sharon E; Mehmeti, Elmira; Anderson, Kevin L; Washburn, Steve P; Baynes, Ronald E

    2016-10-11

    Thymol and carvacrol may be present in several phytoceutical products but there are no well-defined methods to measure these compounds in meat and milk from treated animals. U.S. regulatory authorities deem their presence as an adulteration of food. A rapid and sensitive HS-SPME-GC-MS/MS method was developed for the detection of thymol and carvacrol in bovine milk, plasma, liver, kidney, and fat. Inter- and intraday precision values were all less than 15.7 and 20.2% for thymol and carvacrol, respectively. The accuracy was in ranges of 69.9-111.8% for thymol and 74.0-119.2% for carvacrol. With the exception of fat tissue, stability studies showed that both compounds are stable over a 2 month period. A pilot pharmacokinetic study was conducted to evaluate the developed analytical method and to provide initial estimates of thymol and carvacrol depletion in plasma, milk, and several tissues. Treatment of lactating dairy cattle with phytoceutical products containing these substances resulted in low but measurable residue levels at 96 h for liver and 36 h for milk with very short apparent plasma and milk half-lives (<3.0 h).

  4. Effect of liner design, pulsator setting, and vacuum level on bovine teat tissue changes and milking characteristics as measured by ultrasonography

    PubMed Central

    2004-01-01

    Friesian-type dairy cows were milked with different machine settings to determine the effect of these settings on teat tissue reaction and on milking characteristics. Three teat-cup liner designs were used with varying upper barrel dimensions (wide-bore WB = 31.6 mm; narrow-bore NB = 21.0 mm; narrow-bore NB1 = 25.0 mm). These liners were tested with alternate and simultaneous pulsation patterns, pulsator ratios (60:40 and 67:33) and three system vacuum levels (40, 44 and 50 kPa). Teat tissue was measured using ultrasonography, before milking and directly after milking. The measurements recorded were teat canal length (TCL), teat diameter (TD), cistern diameter (CD) and teat wall thickness (TWT). Teat tissue changes were similar with a system vacuum level of either 50 kPa (mid-level) or 40 kPa (low-level). Widening the liner upper barrel bore dimension from 21.0 mm (P < 0.01) or 25.0 mm (P < 0.001) to 31.6 mm increased the magnitude of changes in TD and TWT after machine milking. Milk yield per cow was significantly (P < 0.05) higher and cluster-on time was reduced (P < 0.01) with the WB cluster as compared to the NB1 cluster. Minimum changes in teat tissue parameters were achieved with system vacuum level of 40 kPa and 50 kPa using NB and WB clusters, respectively. Similar changes in teat tissue and milk yield per cow were observed with alternate and simultaneous pulsation patterns. Widening pulsator ratio from 60:40 to 67:33 did not have negative effects on changes in teat tissue and had a positive effect on milk yield and milking time. Milk liner design had a bigger effect on teat tissue changes and milking characteristics than pulsation settings. PMID:21851658

  5. Effect of liner design, pulsator setting, and vacuum level on bovine teat tissue changes and milking characteristics as measured by ultrasonography.

    PubMed

    Gleeson, David E; O'Callaghan, Edmond J; Rath, Myles V

    2004-05-01

    : Friesian-type dairy cows were milked with different machine settings to determine the effect of these settings on teat tissue reaction and on milking characteristics. Three teat-cup liner designs were used with varying upper barrel dimensions (wide-bore WB = 31.6 mm; narrow-bore NB = 21.0 mm; narrow-bore NB1 = 25.0 mm). These liners were tested with alternate and simultaneous pulsation patterns, pulsator ratios (60:40 and 67:33) and three system vacuum levels (40, 44 and 50 kPa). Teat tissue was measured using ultrasonography, before milking and directly after milking. The measurements recorded were teat canal length (TCL), teat diameter (TD), cistern diameter (CD) and teat wall thickness (TWT).Teat tissue changes were similar with a system vacuum level of either 50 kPa (mid-level) or 40 kPa (low-level). Widening the liner upper barrel bore dimension from 21.0 mm (P < 0.01) or 25.0 mm (P < 0.001) to 31.6 mm increased the magnitude of changes in TD and TWT after machine milking. Milk yield per cow was significantly (P < 0.05) higher and cluster-on time was reduced (P < 0.01) with the WB cluster as compared to the NB1 cluster. Minimum changes in teat tissue parameters were achieved with system vacuum level of 40 kPa and 50 kPa using NB and WB clusters, respectively. Similar changes in teat tissue and milk yield per cow were observed with alternate and simultaneous pulsation patterns. Widening pulsator ratio from 60:40 to 67:33 did not have negative effects on changes in teat tissue and had a positive effect on milk yield and milking time. Milk liner design had a bigger effect on teat tissue changes and milking characteristics than pulsation settings.

  6. Immunohistochemical analysis of cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA) in murine, canine, bovine and equine cerebrospinal tissues.

    PubMed

    Tokunaga, Satoshi; Fujiki, Makoto; Yabuki, Akira; Misumi, Kazuhiro

    2012-04-01

    Cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA), which appears abundantly in hypertrophic cartilage at the stage of endochondral ossification, is also detected in cerebrospinal fluid (CSF) following spinal cord injury. In this study, the localization of the CD-RAP/MIA molecule in normal tissues of the spine and brain obtained from mice, rats, dogs, cattle and horses was examined using immunohistochemistry with a specific antibody. The positive signals of CD-RAP/MIA were found at nerve cells in the spinal cords of all species and were especially strong at cerebellar Purkinje cells. The results suggested that CD-RAP/MIA included in normal cerebrospinal tissues could be a biomarker associated with tissue injuries, as the molecules might flow into the CSF.

  7. Changes in WNT signaling-related gene expression associated with development and cloning in bovine extra-embryonic and endometrial tissues during the peri-implantation period.

    PubMed

    Biase, Fernando H; Rabel, Chanaka; Guillomot, Michel; Sandra, Olivier; Andropolis, Kalista; Olmstead, Colleen; Oliveira, Rosane; Wallace, Richard; Le Bourhis, Daniel; Richard, Christophe; Campion, Evelyne; Chaulot-Talmon, Aurélie; Giraud-Delville, Corinne; Taghouti, Géraldine; Jammes, Hélène; Hue, Isabelle; Renard, Jean Paul; Lewin, Harris A

    2013-12-01

    We determined if somatic cell nuclear transfer (SCNT) cloning is associated with WNT-related gene expression in cattle development, and if the expression of genes in the WNT pathway changes during the peri-implantation period. Extra-embryonic and endometrial tissues were collected at gestation days 18 and 34 (d18, d34). WNT5A, FZD4, FZD5, LRP5, CTNNB1, GNAI2, KDM1A, BCL2L1, and SFRP1 transcripts were localized in extra-embryonic tissue, whereas SFRP1 and DKK1 were localized in the endometrium. There were no differences in the localization of these transcripts in extra-embryonic tissue or endometrium from SCNT or artificial insemination (AI) pregnancies. Expression levels of WNT5A were 11-fold greater in the allantois of SCNT than AI samples. In the trophoblast, expression of WNT5A, FZD5, CTNNB1, and DKK1 increased significantly from d18 to d34, whereas expression of KDM1A and SFRP1 decreased, indicating that implantation is associated with major changes in WNT signaling. SCNT was associated with altered WNT5A expression in trophoblasts, with levels increasing 2.3-fold more in AI than SCNT conceptuses from d18 to d34. In the allantois, expression of WNT5A increased 6.3-fold more in SCNT than AI conceptuses from d18 to d34. Endometrial tissue expression levels of the genes tested did not differ between AI or SCNT pregnancies, although expression of individual genes showed variation across developmental stages. Our results demonstrate that SCNT is associated with altered expression of specific WNT-related genes in extra-embryonic tissue in a time- and tissue-specific manner. The pattern of gene expression in the WNT pathway suggests that noncanonical WNT signal transduction is important for implantation of cattle conceptuses. © 2013 Wiley Periodicals, Inc.

  8. [Unexplained right-sided heart failure following open heart surgery: mind the pericardium].

    PubMed

    Swinkels, B M; Braam, R L; Rensing, B J W M; Jaarsma, W; Defauw, J J A M; Plokker, H W M

    2008-08-30

    Three men, aged 67 years, 80 years and 53 years, respectively, developed signs and symptoms of progressive right-sided heart failure following open heart surgery. They were diagnosed with constrictive pericarditis based on echocardiography, cardiac magnetic resonance and cardiac catheterisation. Following pericardiectomy, two of the patients fully recovered, while one, the 80-year-old man, died during convalescence. When signs and symptoms of progressive right-sided heart failure develop after open heart surgery, a diagnosis of constrictive pericarditis should be considered. Constrictive pericarditis after open heart surgery may be caused by inflammation of the pericardium; an old, fibrotic haemopericardium, which may be diffuse or loculated; pericardial adhesions; or a combination of these entities. Diagnosing constrictive pericarditis is difficult and may take a long time. However, it is important to recognise this disorder early before it has progressed to an advanced stage. Pericardiectomy is the only effective therapy. When performed too late, survival is significantly reduced.

  9. Pulmonary Myofibroblastic Tumour Involving the Pericardium and Left Atrium in an 18 Month Infant.

    PubMed

    Lamas-Pinheiro, Ruben; Rodesch, Gregory; Devalck, Christine; Segers, Valérie; Khelif, Karim; Cappello, Matteo; Steyaert, Henri

    2016-10-20

    Inflammatory myofibroblastic tumor (IMT) is the most frequent primary lung tumor in children and it may be locally aggressive. The management of a locally advanced pulmonary IMT in an 18 month-old female child is presented.A left pulmonary mass was incidentally found on the computerized tomography (CT) scan of a child with persistent systemic inflammatory syndrome. Biopsy confirmed the diagnosis; after preoperative corticotherapy, left pneumonectomy was performed. The pericardium and left atrium were invaded and resected, requiring pericardial reconstruction. There is no relapse at four years of follow-up.Steroids play a role in tumor size reduction, but marginal resection is the gold standard. Extended approaches are feasible and often required in advanced cases.

  10. Genome-wide DNA methylation profiles and their replationship with mRNA and the microRNA transcriptome in bovine muscle tissue (Bos Taurine)

    USDA-ARS?s Scientific Manuscript database

    DNA methylation is a key epigenetic modification in mammals, having essential and important roles in muscle development. We sample longissimus thoracis tissues from a well-known elite native breed of Chinese Qinchuan cattle living within comparable environments at fetal and adult stages, using methy...

  11. Failed detection of Bovine viral diarrhea virus 2 subgenotype a (BVDV-2a) by direct fluorescent antibody test on tissue samples due to reduced reactivity of field isolates to raw anti-BVDV antibody.

    PubMed

    Yan, Lifang; Pace, Lanny W; Baughman, Brittany; Wilson, Floyd D; Zhang, Shuping; Zhang, Michael Z

    2016-03-01

    Bovine viral diarrhea virus 1 (BVDV-1) is associated with mild or subclinical infections, whereas BVDV-2 is frequently implicated in outbreaks of severe thrombocytopenia and acute fatal disease. In the present study, the carcass of a beef breed cow and tissue samples of a beef calf were received for laboratory diagnosis. Both animals exhibited severe clinical signs compatible with thrombocytopenia or hemorrhagic syndrome. Direct fluorescent antibody test (DFAT) failed to detect BVDV antigen in the tissue specimens of both cases. However, immunohistochemistry (IHC) revealed the presence of BVDV antigen in oral and esophageal mucosa and Peyer patches of the beef breed cow. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) detected BVDV-2 in selected tissues of both animals. Subsequently, BVDV was isolated from both cases and subjected to genetic and serologic characterizations. Mutations in the 5'-untranslated genomic region (5'-UTR) primer and probe binding sites and the E2 gene were associated with reduced efficiency of an established real-time RT-PCR assay and amino acid alterations in the E2 glycoprotein, respectively. Both viral isolates were classified by real-time RT-PCR and phylogenetic analysis as BVDV-2 subgenotype a. Unlike BVDV reference strains Singer and 125c, the isolates cross-reacted with anti-BVDV-1 and anti-BVDV-2 reference sera, indicating antigenic variations in field isolates. The isolates also showed reduced reactivity to porcine anti-BVDV antiserum (the raw serum used to produce BVDV DFA conjugate). In summary, data from the present investigation indicated that genetic and antigenic variations affected the performance of detection assays, especially DFAT, highlighting the need for regular evaluation and modification of BVDV tests.

  12. A novel equine-derived pericardium membrane for dural repair: A preliminary, short-term investigation

    PubMed Central

    Centonze, Roberto; Agostini, Emiliano; Massaccesi, Samantha; Toninelli, Stefano; Morabito, Letterio

    2016-01-01

    Background: A large variety of biological and artificial materials are employed in dural repair, each of them with major limitations. Autologous grafts have limited availability and require an additional incision and surgical time. Cadaveric preparations and heterologous materials entail the risk of iatrogenic transmission of prions, whereas synthetic substitutes have been reported to cause inflammatory reactions and graft rejection. An equine-derived pericardium membrane has been developed (Heart®, Bioteck, Vicenza, Italy) with mechanical and safety-related features that could make it suitable for neurosurgical application. Aims: This preliminary study aimed to evaluate the short-term safety and efficacy of the Heart® membrane in dural repair procedures following meningioma surgeries. Subjects and Methods: Medical records of patients who were surgically treated for an intracranial meningioma and underwent duraplasty with the Heart® membrane were reviewed retrospectively. The occurrence of any graft-related complications such as cerebrospinal fluid (CSF) leakage, postoperative hematoma, wound infection, meningitis, and neurological symptoms was analyzed. Results: Eight patients were identified as meeting the inclusion criteria. A watertight closure was achieved in all of them. Postoperatively, no patients exhibited CSF leak, cerebral contusion, hemorrhage, or wound infection. The 1-month radiological follow-up revealed no evidence of pseudomeningocele, wound breakdown, or meningitis. Neurologic complications were observed in three patients but not directly imputable to the dural substitute or its application. Conclusions: In all the patients, the pericardium membrane enabled achievement of a watertight dural closure without graft-related adverse events. Further investigations should be performed to assess medium- and long-term clinical outcomes in a larger set of patients. PMID:27366245

  13. Establishment and characterization of feeder-cell-dependent bovine fetal liver cell lines

    USDA-ARS?s Scientific Manuscript database

    The establishment and initial characterization of bovine fetal liver cell lines is described. Bovine fetal hepatocytes were cultured from the liver of a 34-day bovine fetus by physical disruption of the liver tissue. Released liver cells and clumps of cells were plated on STO feeder layers and wer...

  14. Bovine viral diarrhea virus 1b fetal infection with extensive hemorrhages

    USDA-ARS?s Scientific Manuscript database

    Bovine viral diarrhea virus (BVDV) subtype 1b was isolated from tissues of a term bovine fetus with hemorrhages in multiple tissues. At autopsy, multiple petechial hemorrhages were observed at gross examination throughout the body and placenta. Lung, kidney, thymus, and liver fresh tissues were exam...

  15. Aortic valve replacement with autologous pericardium: long-term follow-up of 15 patients and in vivo histopathological changes of autologous pericardium

    PubMed Central

    Liu, Xiaohong; Han, Lin; Song, Zhigang; Tan, Mengwei; Gong, Dejun; Xu, Zhiyun

    2013-01-01

    OBJECTIVES The study aimed to assess the long-term follow-up of patients with an autologous pericardial aortic valve (APAV) replacement and to analyse in vivo histopathological changes in implanted APAVs. METHODS From 1996 to 1997, 15 patients (mean age, 34 years) underwent aortic valve replacement with the glutaraldehyde-treated autologous pericardium. All patients were followed up after discharge. The excised APAVs were processed for haematoxylin–eosin, Victoria blue-van Gieson and immunohistochemical staining. RESULTS The mean clinical follow-up was 11.43 ± 4.50 years. APAV-related in-hospital and late mortalities were both 0%. Five (33%) patients required reoperation because of a prolapse of the right coronary cusp (n = 1), infective endocarditis (n = 1) or fibrocalcific degeneration (n = 3). Freedom from endocarditis, fibrocalcific degeneration and reoperation at the end of follow-up was 93, 80 and 67%, respectively. The remaining 10 patients were alive and well with a mean New York Heart Association class of 1.10 ± 0.32 and normally functioning aortic valves (peak pressure gradient: 7.70 ± 3.41 mmHg; mean pressure gradient: 1.79 ± 0.64 mmHg). Histopathology revealed that (i) a thin factor VIII-positive layer (endothelialization) was found on all non-endocarditis APAVs; (ii) pericardial cells in all APAVs were positive for α-smooth muscle actin (myofibroblast phenotype) and some cells in the fibrocalcific APAVs were positive for alkaline phosphatase (osteoblast phenotype) and (iii) an elastic band was found in 3 cases (in vivo >9 years). CONCLUSIONS APAV replacement is a procedure with a low mortality. APAVs adapt to new environmental demands by producing an elastic band and by endothelialization, whereas myofibroblast/osteoblast transdifferentiation seems to be responsible for the fibrocalcification of APAVs. PMID:23143205

  16. Aortic valve replacement with autologous pericardium: long-term follow-up of 15 patients and in vivo histopathological changes of autologous pericardium.

    PubMed

    Liu, Xiaohong; Han, Lin; Song, Zhigang; Tan, Mengwei; Gong, Dejun; Xu, Zhiyun

    2013-02-01

    The study aimed to assess the long-term follow-up of patients with an autologous pericardial aortic valve (APAV) replacement and to analyse in vivo histopathological changes in implanted APAVs. From 1996 to 1997, 15 patients (mean age, 34 years) underwent aortic valve replacement with the glutaraldehyde-treated autologous pericardium. All patients were followed up after discharge. The excised APAVs were processed for haematoxylin-eosin, Victoria blue-van Gieson and immunohistochemical staining. The mean clinical follow-up was 11.43 ± 4.50 years. APAV-related in-hospital and late mortalities were both 0%. Five (33%) patients required reoperation because of a prolapse of the right coronary cusp (n = 1), infective endocarditis (n = 1) or fibrocalcific degeneration (n = 3). Freedom from endocarditis, fibrocalcific degeneration and reoperation at the end of follow-up was 93, 80 and 67%, respectively. The remaining 10 patients were alive and well with a mean New York Heart Association class of 1.10 ± 0.32 and normally functioning aortic valves (peak pressure gradient: 7.70 ± 3.41 mmHg; mean pressure gradient: 1.79 ± 0.64 mmHg). Histopathology revealed that (i) a thin factor VIII-positive layer (endothelialization) was found on all non-endocarditis APAVs; (ii) pericardial cells in all APAVs were positive for α-smooth muscle actin (myofibroblast phenotype) and some cells in the fibrocalcific APAVs were positive for alkaline phosphatase (osteoblast phenotype) and (iii) an elastic band was found in 3 cases (in vivo >9 years). APAV replacement is a procedure with a low mortality. APAVs adapt to new environmental demands by producing an elastic band and by endothelialization, whereas myofibroblast/osteoblast transdifferentiation seems to be responsible for the fibrocalcification of APAVs.

  17. Cloning and expression of bovine glucose transporter GLUT12.

    PubMed

    Miller, Peter J; Finucane, Kiera A; Hughes, Megan; Zhao, Feng-Qi

    2005-11-01

    GLUT12 is a new member of facilitative glucose transporters. It was originally cloned from a human breast cancer cell line and its expression has been detected in rat mammary gland. Glucose transport across the plasma membrane of mammary epithelial cells is a rate-limiting factor in milk production. To examine GLUT12's expression and facilitate the study of GLUT12's potential role in supporting milk synthesis in lactating bovine mammary gland, we cloned bovine GLUT12 and examined its distribution of mRNA expression in bovine tissues. The full-length mRNA of bGLUT12 is 2,423 base pairs long and is predicted to encode a protein of 621 amino acids with a molecular weight of approximately 67 kDa. The deduced amino acid sequence of bovine GLUT12 is 87% and 82% identical to the sequences of human and mouse GLUT12. The sequence of bGLUT12 contains several characteristically conserved sugar transporter family signatures. Analysis of current bovine genomic data indicates that bovine GLUT12 gene consists of five exons. The major in vitro transcription and translation product of bovine GLUT12 cDNA migrated at an apparent molecular weight of 41 kDa. In the presence of canine microsomal membranes, the translation product increased to 43 kDa, suggesting glycosylation. GLUT12 mRNA was found in all bovine tissues examined, but most abundant in bovine spleen and skeletal muscle, at intermediate levels in bovine kidney, testes, and mammary gland, and at lower levels in bovine liver, lung and intestine. Immunofluorescence staining showed that, in the presence of insulin, bGLUT12 is mainly distributed in the cytoplasm of the transiently transfected MAC-T bovine mammary epithelial cells.

  18. Effects of nutrient restriction of bovine dams during early gestation on postnatal growth, carcass and organ characteristics, and gene expression in adipose tissue and muscle.

    PubMed

    Long, N M; Prado-Cooper, M J; Krehbiel, C R; DeSilva, U; Wettemann, R P

    2010-10-01

    Angus x Hereford heifers (15 mo and artificially inseminated to a single sire) were used to evaluate the effect of prenatal nutritional restriction on postnatal growth and development. At d 32 of gestation, dams were stratified by BW and BCS and allotted to a low-nutrition [55% of NRC (1996) requirements, n = 10] or moderate-nutrition [100% of NRC (1996) requirements, n = 10] diet. After 83 d of feeding, dams were commingled and received a diet in excess of requirements. Dams were allowed to calve naturally, and birth weights and growth of calves were recorded. Bulls were castrated at birth. Steers (16 mo of age, 5 per treatment) received a high-concentrate diet ad libitum to a constant age (88 ± 1 wk). Steers were slaughtered and weights of the empty body and organs were recorded. Samples of organs, muscle (complexus), and perirenal and subcutaneous adipose tissue were stored at -80 degrees C, and then DNA and protein concentrations were quantified and expression of genes associated with fatty acid metabolism and glucose uptake were measured in adipose and muscle tissue. Dams had similar (P > 0.33) BW and BCS at the beginning of the experiment. At the end of restriction, dams on the low-nutrition diet weighed less (P ≤ 0.01) and had less BCS (P < 0.001) than those on the moderate-nutrition diet. Length of gestation was 274 ± 2 d for dams in the low-nutrition treatment and 278 ± 2 d (P = 0.05) for dams in the moderate-nutrition treatment. Nutrient restriction during gestation did not influence birth weight or postnatal growth of calves. Lungs and trachea of steers whose dams were fed the low-nutrition diet weighed less (P = 0.05) at slaughter than those of steers whose dams were fed the moderate-nutrition diet; weights of other organs were not influenced by treatment. Complexus muscle from steers whose dams were fed the low-nutrition diet had a greater (P = 0.04) concentration of DNA and larger muscle fiber area compared with steers whose dams were fed the

  19. Heidenhain variant of Creutzfeldt–Jakob disease in a patient who had bovine bioprosthetic valve implantation

    PubMed Central

    Hashoul, Jehard; Saliba, Waleed; Bloch, Irina; Jabaly-Habib, Haneen

    2016-01-01

    Creutzfeldt–Jakob disease (CJD) is a rare neurodegenerative disorder characterized by rapidly progressing dementia, general neurologic deterioration, and death. When the leading symptoms are visual disturbances, it is termed as the Heidenhain variant of CJD (HvCJD). CJD was reported following prion-contaminated pericardium transplants but never after bovine bioprosthetic cardiac valve. In this case report, we describe HvCJD in a patient who had a bovine bioprosthetic cardiac valve implant. An 82-year-old-woman was referred to neuro-ophthalmology clinic for unexplained visual loss that started 1 month previously. Medical history included aortic valve replacement with bovine bioprosthetic valve. On examination, best-corrected visual acuity was 20/120 in the right eye and 20/200 in the left eye; otherwise, the eye examination was normal. Humphrey visual fields revealed complete right homonymous hemianopsia. Magnetic resonance imaging (MRI) demonstrated nonspecific white matter changes. A week later, she was hospitalized due to memory impairment; repeated MRI and total body computed tomography scan showed no significant findings. Electroencephalography recordings and extremely elevated cerebrospinal fluid tau protein were compatible with CJD. The patient died 3 weeks later; autopsy was not performed. The patient had HvCJD. Ophthalmologists being first to see these patients should be aware of this diagnosis. Contaminated bovine bioprosthetic valve might be another source for prion disease. Further research is required to establish this issue. PMID:27905341

  20. Discrimination between Synthetically Administered and Endogenous Thiouracil Based on Monitoring of Urine, Muscle, and Thyroid Tissue: An in Vivo Study in Young and Adult Bovines.

    PubMed

    Wauters, J; Van Meulebroek, L; Fichant, E; Delahaut, P; Vanhaecke, L

    2017-08-02

    Thiouracil (TU), synthesized for its thyroid-regulating capacities and alternatively misused in livestock for its weight-gaining effects, is acknowledged to have an endogenous origin. Discrimination between low-level abuse and endogenous occurrence is challenging and unexplored in an experimental setting. Therefore, cows (n = 16) and calves (n = 18) were subjected to a rapeseed-supplemented diet or treated with synthetic TU. Significant higher urinary TU levels were recorded after TU administration (tissue of calves and cows following rapeseed supplementation. The latter concentrations were significantly higher in thyroid tissue of calves (22.9-41.8 μg kg(-1)) and cows (16.9-36.7 μg kg(-1)) after synthetic TU administration. These results strongly point toward thyroid analysis as a discriminatory tool.

  1. Teflon-buttressed sutures plus pericardium patch repair left ventricular rupture caused by radiofrequency catheter ablation: A case report.

    PubMed

    Cao, Hao; Zhang, Qi; He, Yanzhong; Feng, Xiaodong; Liu, Zhongmin

    2016-09-01

    Cardiac rupture often occurs after myocardial infarction or chest trauma with a high mortality rate. However, left ventricular rupture caused by radiofrequency catheter ablation (RFCA) is extremely rare. We describe a case of a 61-year-old male who survived from left ventricular rupture caused by a RFCA procedure for frequent ventricular premature contractions. Surgical exploration with cardiopulmonary bypass (CPB) was performed when the signs of cardiac tamponade developed 7 hours after the ablation surgery. Teflon-buttressed sutures of the tear in the left ventricular posterolateral wall and pericardium patch applied to the contusion region on the wall repaired the rupture safely and effectively. Timely surgical intervention under CPB facilitated the survival of the patient. Teflon-buttressed sutures plus pericardium patch achieved the successful repair of the rupture.

  2. Optimal ratios of essential amino acids stimulate β-casein synthesis via activation of the mammalian target of rapamycin signaling pathway in MAC-T cells and bovine mammary tissue explants.

    PubMed

    Li, S S; Loor, J J; Liu, H Y; Liu, L; Hosseini, A; Zhao, W S; Liu, J X

    2017-08-01

    Amino acids are the building blocks of proteins and serve as key molecular components upstream of the signaling pathways that regulate protein synthesis. The objective of this study was to systematically investigate the effect of essential AA ratios on milk protein synthesis in vitro and to elucidate some of the underlying mechanisms. Triplicate cultures of MAC-T cells and bovine mammary tissue explants (MTE) were incubated with the optimal AA ratio (OPAA; Lys:Met, 2.9:1; Thr:Phe, 1.05:1; Lys:Thr, 1.8:1; Lys:His, 2.38:1; and Lys:Val, 1.23:1) in the presence of rapamycin (control), OPAA, a Lys:Thr ratio of 2.1:1, a Lys:Thr ratio of 1.3:1, a Lys:His ratio of 3.05:1, or a Lys:Val ratio of 1.62:1 for 12 h; the other AA concentrations were equal to OPAA. In some experiments, the cells were cultured with OPAA with or without rapamycin (100 ng/mL) or with mammalian target of rapamycin (mTOR) small interference RNA, and the MTE were exposed to OPAA with rapamycin for β-casein expression. Among the treatments, the expression of β-casein was greatest in the MTE cultured with OPAA. In MAC-T cells, the OPAA upregulated the mRNA expression of SLC1A5 and SLC7A5 but downregulated the expression of IRS1, AKT3, EEF1A1, and EEF2 compared with the control. The OPAA had no effect on the mTOR phosphorylation status but increased the phosphorylation of S6K1 and RPS6. When the MTE were treated with rapamycin in the presence of OPAA, the expression of β-casein was markedly decreased. The phosphorylation of RPS6 and 4EBP1 also was reduced in MAC-T cells. A similar negative effect on the expression of RPS6KB1 and EIF4EBP1 was detected when the cells were cultured with either rapamycin or mTOR small interference RNA. The optimal AA ratio stimulated β-casein expression partly by enhancing the transport of AA into the cells, cross-talk with insulin signaling and a subsequent enhancement of mTOR signaling, or translation elongation in both MAC-T cells and bovine MTE. Copyright © 2017

  3. Bovine coronavirus associated syndromes.

    PubMed

    Boileau, Mélanie J; Kapil, Sanjay

    2010-03-01

    Bovine coronaviruses, like other animal coronaviruses, have a predilection for intestinal and respiratory tracts. The viruses responsible for enteric and respiratory symptoms are closely related antigenically and genetically. Only 4 bovine coronavirus isolates have been completely sequenced and thus, the information about the genetics of the virus is still limited. This article reviews the clinical syndromes associated with bovine coronavirus, including pneumonia in calves and adult cattle, calf diarrhea, and winter dysentery; diagnostic methods; prevention using vaccination; and treatment, with adjunctive immunotherapy.

  4. 78 FR 73993 - Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-10

    ... Health Inspection Service 9 CFR Parts 92, 93, 94, 95, 96, and 98 RIN 0579-AC68 Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products Corrections In rule document 2013-28228 appearing on...

  5. 77 FR 20319 - Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-04

    ...; ] DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service 9 CFR Part 93 RIN 0579-AC68 Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products Correction In proposed rule document...

  6. Deproteinized bovine bone in association with guided tissue regeneration or enamel matrix derivatives procedures in aggressive periodontitis patients: a 1-year retrospective study.

    PubMed

    Artzi, Zvi; Tal, Haim; Platner, Ori; Wasersprung, Nadav; Weinberg, Evgeny; Slutzkey, Shimshon; Gozali, Nir; Carmeli, Guy; Herzberg, Ran; Kozlovsky, Avital

    2015-06-01

    To retrospectively evaluate and compare two regenerative periodontal procedures in young individuals with aggressive periodontitis (AgP). Thirty-two patients aged 14-25 years (mean ± SD 19.3 ± 5.7) were diagnosed as having AgP with multiple intra-bony defects (IBDs) and treated by one of two regenerative modalities of periodontal therapy: guided tissue regeneration (GTR) using deproteinized bone xenograft (DBX) particles and a resorbable membrane (the GTR group), or an application of enamel matrix derivatives (EMD) combined with DBX (the EMD/DBX group). Periodic monitoring of treated sites included recording of probing depth (PD), clinical attachment level (CAL) and gingival recession. Pre-treatment and 1-year post-operative findings were statistically analysed within and between groups. The PD and CAL values decreased significantly with time, but not those between study groups. The mean pre-treatment and 1-year post-treatment PDs of the IBDs of the GTR group (n = 16; sites = 67) were 8.93 ± 1.14 mm and 3.58 ± 0.50 mm, respectively, and the mean CALs were 9.03 ± 1.03 mm and 4.16 ± 0.53 mm respectively. The mean PDs of the EMD/DBX group (n = 16; sites = 73) were 8.77 ± 1.04 mm and 3.61 ± 0.36 mm, respectively, and the mean CALS were 8.79 ± 1.04 mm and 3.77 ± 0.22 mm respectively (p < 0.001 for all). Surgical treatment of AgP patients by either GTR or by application of EMD/DBX yielded similarly successful clinical results at 1-year post-treatment. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Use of Equine Derived Pericardium as a Biological Cover To Promote Closure of a Complicated Wound With Associated Scleroderma and Raynaud's Disease .

    PubMed

    Mulder, Gerit; Lee, Daniel K

    2009-11-01

    A 39-year-old man with previously undiagnosed scleroderma was admitted to the UCSD Medical Center with bilateral, limb-threatening necrotic lower extremity ulcers extending to underlying fascia and muscle. Rather than amputate the extremities, the patient requested alternative treatment and underwent extensive tissue debridement followed by placement of an equine pericardium xenograft. Subsequent to treatment, the patient underwent weekly examinations and dressing changes without additional treatment. The patient was ambulating without assistance and with complete closure of all wounds in 10 weeks. The patient remained without wound recurrence at a recent 6-month follow-up visit .

  8. Unlocking the bovine genome

    USDA-ARS?s Scientific Manuscript database

    The draft genome sequence of cattle (Bos taurus) has now been analyzed by the Bovine Genome Sequencing and Analysis Consortium and the Bovine HapMap Consortium, which together represent an extensive collaboration involving more than 300 scientists from 25 different countries. ...

  9. Bovine adipose triglyceride lipase is not altered and adipocyte fatty acid binding protein is increased by dietary flaxseed

    USDA-ARS?s Scientific Manuscript database

    In this paper, we report the full length coding sequence of bovine ATGL cDNA are reported and analyze its expression in bovine tissues. Similar to human, mouse, and pig ATGL sequences, bovine ATGL has a highly conserved patatin domain that is necessary for lipolytic function in mice and humans. Thi...

  10. Description and analysis of the Bovine Gene Atlas - An extensive compendium of bovine transcript profiles

    USDA-ARS?s Scientific Manuscript database

    The Bovine Gene Atlas (BGA) is a compendium of over 7.2 million unique 20-base transcript tags profiled from 81 tissues acquired from the cow “L1 Dominette 01449” (L1D), her male fetus, her 255-day-old heifer calf, and her father. The BGA tags were generated on a next-generation massively parallel ...

  11. Alcohol and the Heart: A Proteomics Analysis of Pericardium and Myocardium in a Swine Model of Myocardial Ischemia.

    PubMed

    Elmadhun, Nassrene Y; Sadek, Ahmed A; Sabe, Ashraf A; Lassaletta, Antonio D; Sellke, Frank W

    2015-11-01

    Previous studies have demonstrated that moderate alcohol consumption is cardioprotective and reduces postoperative pericardial adhesions; however, the mechanism is not fully understood. Using proteomic analysis, we sought to objectively investigate the effects of daily moderate alcohol consumption in the pericardium and myocardium in a swine model of chronic myocardial ischemia. Fourteen swine underwent placement of an ameroid constrictor to induce chronic myocardial ischemia. Animals were supplemented with 90 mL of ethanol daily (ETOH) or 80 g of sucrose of equal caloric value (SUC). After 7 weeks, the ischemic myocardium and pericardium were harvested for proteomics analysis. Pericardial proteomics analysis yielded 397 proteins, of which 23 were unique to SUC and 52 were unique to ETOH. Of the 322 common proteins, 71 were statistically significant and 23 were characterized (p < 0.05). Alcohol supplementation increased structural proteins, and decreased immune protease inhibitors and coagulation proteins in the pericardium (p < 0.01). Myocardial proteomics analysis yielded 576 proteins, of which 32 were unique to SUC and 21 were unique to ETOH. Of the 523 common proteins, 85 were significant, and 32 were characterized (p < 0.05). Alcohol supplementation decreased cardiac remodeling proteins, cell death proteins and motor proteins, and increased metabolic proteins (p < 0.05). The results suggest that daily moderate alcohol consumption affects numerous pathways that contribute to cardioprotection, including cardiac remodeling, metabolism, and cell death. Our findings reveal the biosignature of myocardial and pericardial protein expression in the setting of chronic myocardial ischemia and daily moderate alcohol consumption. Copyright © 2015 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.

  12. The visceral pericardium: macromolecular structure and contribution to passive mechanical properties of the left ventricle

    PubMed Central

    Jöbsis, Paul D.; Ashikaga, Hiroshi; Wen, Han; Rothstein, Emily C.; Horvath, Keith A.; McVeigh, Elliot R.; Balaban, Robert S.

    2010-01-01

    Much attention has been focused on the passive mechanical properties of the myocardium, which determines left ventricular (LV) diastolic mechanics, but the significance of the visceral pericardium (VP) has not been extensively studied. A unique en face three-dimensional volumetric view of the porcine VP was obtained using two-photon excitation fluorescence to detect elastin and backscattered second harmonic generation to detect collagen, in addition to standard light microscopy with histological staining. Below a layer of mesothelial cells, collagen and elastin fibers, extending several millimeters, form several distinct layers. The configuration of the collagen and elastin layers as well as the location of the VP at the epicardium providing a geometric advantage led to the hypothesis that VP mechanical properties play a role in the residual stress and passive stiffness of the heart. The removal of the VP by blunt dissection from porcine LV slices changed the opening angle from 53.3 ± 10.3 to 27.3 ± 5.7° (means ± SD, P < 0.05, n = 4). In four porcine hearts where the VP was surgically disrupted, a significant decrease in opening angle was found (35.5 ± 4.0°) as well as a rightward shift in the ex vivo pressure-volume relationship before and after disruption and a decrease in LV passive stiffness at lower LV volumes (P < 0.05). These data demonstrate the significant and previously unreported role that the VP plays in the residual stress and passive stiffness of the heart. Alterations in this layer may occur in various disease states that effect diastolic function. PMID:17933976

  13. Aortic valve reconstruction using autologous pericardium for ages over 80 years.

    PubMed

    Ozaki, Shigeyuki; Kawase, Isamu; Yamashita, Hiromasa; Uchida, Shin; Nozawa, Yukinari; Takatoh, Mikio; Hagiwara, So; Kiyohara, Nagaki

    2014-10-01

    We performed original aortic valve reconstruction using autologous pericardium; the feasibility for elderly patients is reviewed. From April 2007 through September 2011, aortic valve reconstruction was carried out in 86 patients over the age of 80 years. Twenty-seven patients were male and 59 were female. Mean age was 82.9 ± 2.5 years. Seventy-two patients had aortic stenosis and 14 had aortic regurgitation. Mean preoperative surgical annular diameter was 19.5 ± 2.5 mm. There were 80 (90.7%) cases of small aortic annulus. Mean preoperative logistic EuroSCORE was 22.9 ± 15.8. Isolated aortic valve reconstructions were performed in 51 patients. Concomitant procedures included coronary artery bypass grafting in 6, hemiarch aortic replacements in 6, 9 maze procedures, and some combinations. No conversion to valve replacement was required. Mean follow-up was 1243 days. There were 3 hospital deaths due to noncardiac causes. No reoperation was needed. Survival at 56 months was 87.0%. No thromboembolic event occurred. Echocardiography 3.5 years after surgery revealed an average peak pressure gradient of 14.6 ± 3.8 mm Hg. No moderate or severe regurgitation was recorded. Aortic valve reconstruction is feasible for patients older than 80 years, resulting in good hemodynamics and a better quality of life, without anticoagulation. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  14. Surgical reconstruction of the left main coronary artery: fresh autologous pericardium or saphenous vein patch.

    PubMed

    Raanani, Ehud; Kogan, Alexander; Shapira, Yaron; Sagie, Alex; Kornowsky, Ran; Vidne, Bernardo A

    2004-11-01

    Isolated stenosis of the left main coronary artery is usually treated by coronary bypass surgery. However, this approach restores a less physiologic perfusion of the myocardium that leads to occlusion of the left main coronary artery, and restores only a retrograde perfusion of a rather extensive myocardial area. Coronary ostial plasty has been described as an alternative surgical technique in isolated ostial left main coronary artery stenosis without calcification. We review our experience with 15 patients. After placing the patient on cardiopulmonary bypass and aortic cross clamping, the main pulmonary trunk was retracted laterally. The left main coronary artery was approached anteriorly through a curved aortotomy. Reconstruction was performed using fresh pericardial patch or saphenous vein that was tailored as a patch. There were no early mortality or perioperative myocardial infarctions. During mean follow-up of 55 +/- 39 months, no patients had any cardiac events or required repeated coronary intervention. All patients underwent follow-up transesophageal echocardiography, which demonstrated a wide open left main coronary artery (range 3 to 8 mm), normal flow pattern by pulsed-wave Doppler, and no aneurysmal dilatation or calcification of the onlay patch. Coronary angiography and intravascular ultrasound were performed in 2 patients because of nonspecific chest discomfort. In both cases, the left main coronary artery was found to be wide open, and there were no signs of patch calcification. Surgical reconstruction of the left main coronary artery is safe and effective for the treatment of selected cases of isolated left main stenosis. The use of autologous pericardium appears to be as safe as saphenous vein patch. Neither method was associated with postoperative aneurysmal dilatation or calcification.

  15. Developmental abnormalities in mice transgenic for bovine oncostatin M.

    PubMed Central

    Malik, N; Haugen, H S; Modrell, B; Shoyab, M; Clegg, C H

    1995-01-01

    Oncostatin M belongs to the subfamily of hematopoietin cytokines that binds a receptor complex containing gp130. To date, only the human form of oncostatin M has been identified, and its evolutionary conservation is unresolved. We have isolated a bovine gene whose open reading frame encodes a precursor protein that is 58% identical to human oncostatin M. A comparison of the bovine and human amino acid sequences predicts significant similarity, including the four-alpha-helical-bundle structure and the placement of disulfide bridges. As with the human protein, bovine oncostatin M binds specific receptors on human H2981 cells and inhibits the proliferation of human A375 tumor cells and mouse M1 leukemia cells. To identify activities regulated in vivo, we injected bovine oncostatin M fusion genes containing various tissue-specific promoters into mouse embryos. The frequencies of transgenic mice were reduced significantly, suggesting that overexpression of the bovine cytokine is detrimental to normal mouse development. In addition to deaths associated with expression in neurons and keratinized epithelia, bovine oncostatin M caused abnormalities in bone growth and spermatogenesis, stimulated fibrosis surrounding islets in the pancreas, and disrupted normal lymphoid tissue development. This work establishes the existence of a nonprimate oncostatin M gene and provides the first demonstration that this cytokine can function in a pleiotropic manner in vivo. Information regarding bovine oncostatin M may help characterize the structure and function of this cytokine in other vertebrate species. PMID:7739518

  16. Bovine dedifferentiated adipose tissue (DFAT) cells

    PubMed Central

    Wei, Shengjuan; Du, Min; Jiang, Zhihua; Duarte, Marcio S; Fernyhough-Culver, Melinda; Albrecht, Elke; Will, Katja; Zan, Linsen; Hausman, Gary J; Elabd, Elham M Youssef; Bergen, Werner G; Basu, Urmila; Dodson, Michael V

    2013-01-01

    Dedifferentiated fat cells (DFAT cells) are derived from lipid-containing (mature) adipocytes, which possess the ability to symmetrically or asymmetrically proliferate, replicate, and redifferentiate/transdifferentiate. Robust cell isolation and downstream culture methods are needed to isolate large numbers of DFAT cells from any (one) adipose depot in order to establish population dynamics and regulation of the cells within and across laboratories. In order to establish more consistent/repeatable methodology here we report on two different methods to establish viable DFAT cell cultures: both traditional cell culture flasks and non-traditional (flat) cell culture plates were used for ceiling culture establishment. Adipocytes (maternal cells of the DFAT cells) were easier to remove from flat culture plates than flasks and the flat plates also allowed cloning rings to be utilized for cell/cell population isolation. While additional aspects of usage of flat-bottomed cell culture plates may yet need to be optimized by definition of optimum bio-coating to enhance cell attachment, utilization of flat plate approaches will allow more efficient study of the dedifferentiation process or the DFAT progeny cells. To extend our preliminary observations, dedifferentiation of Wagyu intramuscular fat (IMF)-derived mature adipocytes and redifferentiation ability of DFAT cells utilizing the aforementioned isolation protocols were examined in traditional basal media/differentiation induction media (DMI) containing adipogenic inducement reagents. In the absence of treatment approximately 10% isolated Wagyu IMF-mature adipocytes dedifferentiated spontaneously and 70% DFAT cells displayed protracted adipogenesis 12 d after confluence in vitro. Lipid-free intracellular vesicles in the cytoplasm (vesicles possessing an intact membrane but with no any observable or stainable lipid inside) were observed during redifferentiation. One to 30% DFAT cells redifferentiated into lipid-assimilating adipocytes in the DMI media, with distinct lipid-droplets in the cytoplasm and with no observable lipid-free vesicles inside. Moreover, a high confluence level promoted the redifferentiation efficiency of DFAT cells. Wagyu IMF dedifferentiated DFAT cells exhibited unique adipogenesis modes in vitro, revealing a useful cell model for studying adipogenesis and lipid metabolism. PMID:23991361

  17. Towards evaluating post-irradiation tissue alterations

    NASA Astrophysics Data System (ADS)

    Daar, Eman; Bradley, D. A.; Alkhorayef, M.; Al-Mugren, K. S.; Abdallat, R. G.; Al-Dousari, H.

    2017-08-01

    There is apaucity of data concerning irradiation effects on the extracellular matrix and on organised tissues. Examples of such research are cited as are some of the limiting factors towards obtaining meaningful results. This would engender a range of research towards further improving the quality of life, most pointedly of those receiving radiotherapy. As cancer survivor rates increase, survivors are more likely to experience side effects of radiotherapy. This study examines the effects of radiotherapy doses on the extracellular matrix as hyaluronic acid (HA) and pericardium.

  18. Bovine somatic cell nuclear transfer.

    PubMed

    Ross, Pablo J; Cibelli, Jose B

    2010-01-01

    Somatic cell nuclear transfer (SCNT) is a technique by which the nucleus of a differentiated cell is introduced into an oocyte from which its genetic material has been removed by a process called enucleation. In mammals, the reconstructed embryo is artificially induced to initiate embryonic development (activation). The oocyte turns the somatic cell nucleus into an embryonic nucleus. This process is called nuclear reprogramming and involves an important change of cell fate, by which the somatic cell nucleus becomes capable of generating all the cell types required for the formation of a new individual, including extraembryonic tissues. Therefore, after transfer of a cloned embryo to a surrogate mother, an offspring genetically identical to the animal from which the somatic cells where isolated, is born. Cloning by nuclear transfer has potential applications in agriculture and biomedicine, but is limited by low efficiency. Cattle were the second mammalian species to be cloned after Dolly the sheep, and it is probably the most widely used species for SCNT experiments. This is, in part due to the high availability of bovine oocytes and the relatively higher efficiency levels usually obtained in cattle. Given the wide utilization of this species for cloning, several alternatives to this basic protocol can be found in the literature. Here we describe a basic protocol for bovine SCNT currently being used in our laboratory, which is amenable for the use of the nuclear transplantation technique for research or commercial purposes.

  19. Adipogenesis of bovine perimuscular preadipocytes

    SciTech Connect

    Taniguchi, Masaaki; Le Luo Guan; Zhang Bing; Dodson, Michael V.; Okine, Erasmus; Moore, Stephen S.

    2008-02-01

    In this study, non-transformed progeny adipofibroblasts, derived from mature adipocyte dedifferentiation, was used as a novel in vitro model to study adipogenic gene expression in cattle. Adipofibroblasts from dedifferentiated mature perimuscular fat (PMF) tissue were cultured with differentiation stimulants until the cells exhibited morphological differentiation. Treated cells were harvested from day 2 to 16 for RNA extraction, whereas control cells were cultured without addition of stimulants. Results from time course gene expression assays by quantitative real-time PCR revealed that peroxisome proliferator-activated receptor gamma (PPAR-{gamma}), sterol regulatory element binding protein 1 (SREBP-1) and their six down-stream genes were co-expressed at day 2 post-differentiation induction. When compared to other adipogenesis culture systems, the adipogenic gene expression of bovine PMF adipofibroblasts culture was different, especially to the rodent model. Collectively, these results demonstrated PPAR-{gamma} and SREBP-1 cooperatively play a key role to regulate the re-differentiation of bovine adipofibroblasts, during early conversion stages in vitro.

  20. Tricuspid Anterior Leaflet Replacement with Autologous Pericardium and Polytetrafluoroethylene Chordae, Followed by Edge-to-Edge Repair

    PubMed Central

    García-Rinaldi, Raúl

    2007-01-01

    Treatment of endocarditis of the tricuspid valve is very complex. Valvectomy without replacement can result in significant postoperative morbidity, and replacement is hazardous due to the possibility of prosthetic infection. Repair of the tricuspid valve is the best alternative. Many ingenious methods have been devised to repair the tricuspid valve in patients with infective endocarditis. However, repair of the endocarditic anterior leaflet presents a difficult surgical problem. We have developed a technique to replace the central portion of the anterior leaflet, in which most of the leaflet is excised and replaced with autologous pericardium. By preserving a small rim of the leaflet, we preserve the lateral and medial chordal attachments. Supplemental neochordae are created with polytetrafluoroethylene sutures. An edge-to-edge repair is used to correct residual insufficiency of the valve. We have used this technique in 4 patients. One patient died 1 month after surgery, but with a competent tricuspid valve. Three patients are now alive, 48, 34, and 1.5 months after surgery. Their tricuspid valves are functional, moving well, and competent. We conclude that the centrally infected anterior leaflet of the tricuspid valve can be replaced with autologous pericardium. The implantation of polytetrafluoroethylene neochordae and edge-to-edge repair can be used to achieve valve competence. Short- and medium-term follow-up in patients who have been treated with these techniques demonstrates that the valve remains fully mobile and competent. PMID:17948081

  1. Bovine trophectoderm cells induced from bovine fibroblasts with induced pluripotent stem cell reprogramming factors.

    PubMed

    Talbot, Neil C; Sparks, Wendy O; Phillips, Caitlin E; Ealy, Alan D; Powell, Anne M; Caperna, Thomas J; Garrett, Wesley M; Donovan, David M; Blomberg, Le Ann

    2017-06-01

    Thirteen independent induced bovine trophectroderm (iBT) cell lines were established by reprogramming bovine fetal liver-derived fibroblasts after viral-vector transduction with either six or eight factors, including POU5F1 (OCT4), KLF4, SOX2, MYC, NANOG, LIN28, SV40 large T antigen, and hTERT. Light- and electron-microscopy analysis showed that the iBT cells had epithelial cell morphology typical of bovine trophectoderm cells. Reverse-transcription-PCR assays indicated that all of the cell lines expressed interferon-tau (IFNT) at passages 1 or 2. At later passages (≥ passage 8), however, immunoblot and antiviral activity assays revealed that more than half of the iBT cell lines had stopped expressing IFNT. Messenger RNAs specific to trophectoderm differentiation and function were found in the iBT cell lines, and 2-dimensional-gel analysis for cellular proteins showed an expression pattern similar to that of trophectoderm cell lines derived from bovine blastocysts. Integration of some of the human reprogramming factors, including POU5F1, KLF4, SOX2, MYC, NANOG, and LIN28, were detected by PCR, but their transcription was mostly absent in the iBT cell lines. Gene expression assessment of endogenous bovine reprogramming factor orthologs revealed endogenous bLIN28 and bMYC transcripts in all; bSOX2 and bNANOG in none; and bKLF4 and bPOU5F1 in less than half of the iBT cell lines. These results demonstrate that bovine trophectoderm can be induced via reprogramming factor expression from bovine liver-derived fibroblasts, although other fibroblast populations-e.g., derived from fetal thigh tissue-may produce similar results, albeit at lower frequencies. © 2017 Wiley Periodicals, Inc.

  2. Radiodensity and hardness of enamel and dentin of human and bovine teeth, varying bovine teeth age.

    PubMed

    Fonseca, R B; Haiter-Neto, F; Carlo, H L; Soares, C J; Sinhoreti,