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Perfused vessel density measured by Side stream dark field imaging optical probe Microscan; MicroVision Medical, Amsterdam, Netherlands. Serum potassium measured in milligrams per deciliter. Eligibility Criteria. Inclusion Criteria: Septic shock patients. Aged above 18 years. On high dose vasopressors defined as norepinephrine infusion above 0.

Impaired cardiac contractility Bronchospasm. Major burns Liver failure.


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Active bleeding. Contacts and Locations. Information from the National Library of Medicine To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor. Please refer to this study by its ClinicalTrials. More Information.

National Library of Medicine U. National Institutes of Health U. Department of Health and Human Services.

Methods and Protocols

The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Septic Shock. Phase 3. Study Type :. Estimated Enrollment :. Estimated Study Start Date :. Estimated Primary Completion Date :.


  1. Methods in molecular medicine: angiotensin protocols.
  2. Angiotensin Protocols (Methods in Molecular Medicine).
  3. Global Food Projections to 2020: Emerging Trends and Alternative Futures.
  4. Table of contents.
  5. Estimated Study Completion Date :. Drug: Norepinephrine patients will receive norepinephrine infusion adjusted according to blood pressure Other Name: Noradrenaline. Placebo Comparator: Normal saline group Patients will receive normal saline infusion in addition to norepinephrine infusion. Although the precise mechanism by which STS depresses the development of cardiac fibrosis remains to be further clarified, understanding the pharmacologic actions of STS on cardiac fibroblasts may contribute to the choice of STS for the people who suffer from cardiovascular diseases.

    Peptidomimetics Protocols Methods in Molecular Medicine

    STS All other chemicals used were of the highest grade available commercially. The molecule structure of STS is shown in Figure 1.

    Rat cardiac fibroblasts were prepared and cultured as described earlier. Briefly, cardiac ventricles from one-day-old Wistar rats were separated and minced with scissors into small pieces in ice-cold balanced salt solution.

    The digestion steps were repeated five to seven times until the tissues were completely digested. The cells were then combined, centrifuged, and re-suspended in a chilled fetal calf serum. The supernatant was discarded, and the cells were re-suspended in DMEM. After removal of the myocyte-enriched medium, DMEM was then added to the pre-plated CFs which were cultured for 2 days before being passaged.

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    Experiments were performed with cells from passage 3. Fibroblasts from each group were pelleted and extracted in iced cell lysis buffer Cell Signaling Technologies. The membranes were detected with the ECL system Amersham Life Sciences and relative intensities of protein bands analyzed by Scan-gel-it software. Active MMP-1 secreted into culture medium can be identified and quantified through gelatin zymography. The gels were soaked in 2. The gels were then incubated in a developing buffer containing 50 mM Tris, pH 7.

    After incubation, the gels were stained with Coomassie blue and photographed. After 24 h, cell proliferation was assessed by the MTT assay. The assay is based on the transformation of the tetrazolium salt MTT by active mitochondria to an insoluble formazan salt.

    Bibliographic Information

    DNA synthesis was evaluated by measuring [ 3 H]thymidine incorporation and collagen synthesis was evaluated by measuring [ 3 H]proline incorporation as described earlier Zhang et al. In brief, cardiac fibroblasts were made quiescent by culture in serum-free DMEM for 24 h. The cells were solubilized and the cell extracts analyzed in a liquid scintillation counter. After 12 h, the cells were washed twice with PBS and detached by trypsin, then measured for DCF fluorescence intensity by fluorospectrophotometer analysis at an excitation wavelength of nm and at an emission wavelength of nm.

    The cell number in each sample was counted and utilized to normalize the fluorescence intensity of DCF. Cytochrome c reduction was measured by reading absorbance at nm on a microplate reader. Superoxide production in nanomoles per milligram of protein was calculated from the difference between absorbance with and without SOD and extinction coefficient for change of ferricytochrome c to ferrocytochrome c. Osteopontin is produced by rat cardiac fibroblasts and mediates A II -induced DNA synthesis and collagen gel contraction.

    J Clin Invest ; 98 : - Collagen metabolism in cultured adult rat cardiac fibroblasts: response to angiotensin II and aldosterone. J Mol Cell Cardiol ; 26 : - Brown L. Cardiac extracellular matrix: a dynamic entity. Cytokines regulate matrix metalloproteinases and migration in cardiac fibroblasts. Biochem Biophys Res Commun ; : - Chen J, Mehta JL. Angiotensin II-mediated oxidative stress and procollagen-1 expression in cardiac fibroblasts: blockade by pravastatin and pioglitazone.

    Hypertension a ; 44 : - J Cardiovasc Pharmacol b ; 44 : - Transforming growth factor beta receptor endoglin is expressed in cardiacfibroblasts and modulates profibrogenic actions of angiotensin II. Circ Res c ; 95 : - Pharmacological studies of sodium tanshinone IIA sulfonate. Acta Pharmacol Sin ; 14 : - Cardiac remodeling-concepts and clinical implications: a consensus paper from an international forum on cardiac remodeling. Behalf on an international forum on cardiac remodelling.

    J Am Coll Cardiol ; 35 : - D'Armiento J. Matrix metalloproteinase disruption of the extracellular matrix and cardiac dysfunction. Trends Cardiovasc Med ; 12 : 97 - Cardiac remodeling and failure: From molecules to man Part II. Cardiovasc Pathol ; 14 : 49 - Gibbons GH.

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    The Renin-Angiotensin-Aldosterone System - Methods and Protocols | Sean E. Thatcher | Springer

    The pathophysiology of hypertension: the importance of angiotensin II in cardiovascular remodeling. Am J Hypertens ; 11 : S - S. The dynamic extracellular matrix: intervention strategies during heart failure and atherosclerosis. J Pathol ; : - Distinct upregulation of extracellular matrix genes in transition from hypertrophy to hypertensive heart failure. Hypertension ; 45 : - Inhibition of interleukin and interferon-gamma production in immune cells by tanshinones from Salvia miltiorrhiza.

    Immunopharmacology ; 49 : - What mechanisms underlie diastolic dysfunction in heart failure? Circ Res ; 94 : - Angiotensin II has multiple profibrotic effects in human cardiac fibroblasts. Circulation ; : - Angiotensin II induces cardiac phenotypic modulation and remodeling in vivo in rats. Hypertension ; 25 : - Angiotensin- : an active member of the renin-angiotensin system.

    J Physiol Pharmacol. HOPE Investigators. Counterregulatory actions of angiotensin- Angiotensin- : an update. Regul Pept. Angiotensin- and its receptor as a potential targets for new cardiovascular drugs. Expert Opin Investig Drugs.

    Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors

    Cardiovasc Drug Rev. Antithrombotic effect of captopril and losartan is mediated by angiotensin- Angiotensin- contributes to the antihypertensive effects of blockade of the renin-angiotensin system. The antithrombotic effect of angiotensin- involves mas- mediated NO release from platelets. Mol Med. Angiotensin- is an endogenous ligand for the G protein- coupled receptor Mas. Uekama K. Design and evaluation of cyclodextrin-based drug formulation.