In this third meeting we started working, using a standard blood sample (without disease). To teach the technique, Adriana preparing a tube and we repeated it in triplicate. Part One:
eppendorf tubes put in 500 ul of lysis solution. Add 300 ul of whole blood, taking care not to be formed bubbles. Propipeta mix for evacuation.
add 100 ul of 33% sucrose solution at the base of each tube without mixing the phases. We plugged the tubes and centrifuged at 13500 rpm for 2 minutes.
discarded the supernatant taking care not to remove the pellets.
We wash with 300 ul of PBS, mixing again for evacuation pipette.
add 100 ul of 11% sucrose solution at the base of each tube without mixing the phases. We plugged and re-centrifuge at 13500 rpm for 2 minutes. Again
discard the supernatant taking care not to remove the pellets.
add 75 ul of NaOH and vortex.
incubated in the thermocycler at 95 ° C for 14 minutes and 30 ° C for 1 minute. Centifugamos
briefly to lower condensation on the walls of the tube (use the "press").
neutralized with 10.5 ul of Tris / HCl and centrifuged 4 minutes at 14000 rpm.
DNA is found in the supernatant. * Note: Each time a tip comes in contact with the sample should be excluded to avoid contaminating the solutions. Since we were using blood from one person, we put the necessary solution to each tube (without touching the sample) and after evacuation were mixing by pipette. When working with samples of different people we are going to use a tip for each sample. Part Two: We
DNA to the third floor of the Academy, where was the spectrophotometer. We use a quartz cell plastic because the cells do not efficiently transmit ultraviolet light. We measured the absorbance at 230 nm, 260 nm (maximum absorption) and 280 nm. At the conclusion of the pilot, Dr. Woods continued with the theoretical explanation about what they will do next week (PCR). Then he showed us the program Oligo, used to design primers that can identify potential problems and disadvantages of the design (the union of the chain, annealing temperature of melting, content G and C bases) for the best first time possible to make it.
today Photos: http://www.slide.com/r/IAbJpoLCwz-wsZcy8VSv8bYoiDEOg0_P?previous_view=mscd_embedded_url&view=original
Thursday 29 May Thursday 22 May Thursday 
In this second meeting the doctor continued with the theoretical explanation of Von Willebrand disease. He showed us the different types of tests done for them: bleeding time, ELISA (photo), etc. By Adriana scheme explained how hemophilia is inherited; when one is a carrier (only females), when one has the disease (men only) and cases of healthy men. Finally we saw the different types of VWD (type 1, type 2A, 2B, 2M, 2N and type 3), what distinguishes them and what tests are implemented for a diagnosis. All these cases were exemplified to see what range of data the patient has von Willebrand disease.
May 15 Today we started the project. At this first meeting we knew the place: the library, the area of \u200b\u200bpatient care, laboratories, address ...
Then Dr. Adriana Woods, a researcher with whom we work throughout the year, taught us about Von Willebrand disease (VWD) and hemophilia. We learned the coagulation cascade, laboratory methods to determine the qualitative and quantitative deficiencies of the various factors and interactions of Von Willebrand factor (VWF) to platelets, factor VIII and subendothelial tissue. today Photos: http://www.slide.com/r/kkyBq6nQ2j9kdVRyYkXMTANMgf1Bz1E3?previous_view=mscd_embedded_url&view=original
Curriculum Dr. Adriana Woods CURRICULUM VITAE
ADRIANA INES WOODS
PERSONAL DATA NAME: Adriana Inés Woods
PLACE AND DATE OF BIRTH: Maciel (Santa Fe)
05/31/1953
DOCUMENT: ID: 10,761,901
CI: 11,265,898 (Federal Police)
MARITAL STATUS: Married
ADDRESS: 2367 7th Lacroze Federico B. Federal Capital. (CP: 1426)
PHONE: 4-772-2540
TITLE: Biochemistry (Universidad Nacional de Córdoba) 11/06/1977
CURRENT POSITION: Part of the Support Staff Career of CONICET.
Category: Home Professional.
WORKPLACE: Department of Haemostasis and Thrombosis, Hematological Research Institute 'Mariano R Castex ", National Academy of Medicine.
Pacheco de Melo 3081, Buenos Aires.
WORK HISTORY IN THE CONICET
- Part of the Career of CONICET Support Staff.
categories: professional technical assistant from 03/10/1983 until 31/07/1991.
leading professional technical
from 01/08/1991 to present. WORK HISTORY UNDERGRADUATE
- Practitioner honorary Laboratory Neonatology, Provincial Maternity Hospital, Cordoba, the 06/18/1973 to 31/03/1975.
- Practitioner honorary Central Laboratory, Hospital Nacional de Clínicas "Dr. Peter Vella, a city of Cordoba, from 08/04/1975 to 02/01/1976.
- Practitioner lower Central Laboratory, Hospital Nacional de Clínicas "Dr. Peter Vella, Cordoba DM by Resolution No. 1904 of the Medical Faculty of the UNC, the 01/02/1976 to 31-12 - 76.
- Practitioner greater Central Laboratory, Hospital Nacional de Clínicas "Dr. Peter Vella, a city of Cordoba by Resolution No. of the School of Chemistry, UNC, from 01/01/1977 to 06/30/1977.
- Guard Officer of the Council of Central Laboratory, the National Clinical Hospital "Dr. Peter Vella, a city of Cordoba, from 03/01/1977 to 07/31/1977.
- Practitioner's largest Clinical Laboratory, Department of Clinical Ophthalmology, National Clinical Hospital "Dr. Peter Vella, a city of Cordoba, from 01/02/1977 to 31/10/1977.
UPDATE COURSE GRADE PRE
- Immune Response. Biochemistry Association of Cordoba, in September. 1975.
- Hematology. School of Medical Assistants. Faculty of Medical Sciences, UNC, oct. 1975.
- Topics of Parasitology and Mycology. Practitioners Ateneo Biochemistry. Academic Unit of Medicine, Hospital San Roque, Córdoba, 17 to 21/11/1975.
- Endocrine Laboratory in Obstetrics. Biochemistry Association of Cordoba, and 1/2-9-76 24/25-9-76.
- Sixth Form of hematology, immunology and laboratory. Hemotherapy and Blood Institute. National Clinical Hospital. UNC 11 to 16/10/1976.
- Diabetes. UniMediCor, UNC, 24 to 29/10/1976.
- Chagas Mazza. Biochemistry Association of Cordoba 5 / 6 and 12/13-8-77.
- Renal Transplant and Hemodialysis. Medical Center Kidney Disease and Hypertension.
Córdoba, 08/20/1977.
- Hematology, Immunohematology and Hemotherapy. Society of Hematology and
Hemotherapy of Córdoba. from 22 to 26/08/1977.
- Roundtable on "Behavior of Medicine, Department of Medical Education
Roemmers Laboratories. Cordoba, in September. 1977.
- XIV Course immunopathology. Chair and Institute of Allergy and Immunopathology,
National Hospital de Clinicas, Córdoba. from 08/07/1977 to 09/10/1977.
- Assistant to the Conference and I VIII Argentine Congress of Mycology. Córdoba. from 4 to 8/10/1977.
- Mycological Diagnosis and Immune. Conferences and I VIII Argentine Congress of Mycology. Cordoba
5 to 10/07/1977. GRANTS COMPETITIONS
- FECIC (Foundation Educational, Scientific and Cultural Organization) of 01/05/1978 to 05/01/1980.
Labour Issues:
a) Comparison of a human brain cephalin and commercial phospholipid in the implementation of Activated Partial Thromboplastin Time.
b) Standardization of the technique of Platelet Factor 3 availability.
c) Controls on Antithrombotic Therapy. Presented at the Sociedad Argentina de Investigación Clínica, Mar del Plata, November 1979.
d) Detection Technique platelet microaggregates: an attempt at clinical application.
Workplace: Department of Haemostasis and Thrombosis, Hematological Research Institute, National Academy of Medicine.
- CONICET, 04/01/1980 to 01/04/1982.
Category: Initiation. Theme
work: Participation of platelets in the fibrinolytic system.
Scholarship Director: Dr Maria Angela Lazzari Lopez.
Workplace: Department of Haemostasis and Thrombosis, Hematological Research Institute 'Mariano R. Castex ", National Academy of Medicine. Buenos Aires.
PARTICIPATION IN CONFERENCES AND SCIENTIFIC MEETINGS
- Days of arterial and venous thrombosis. Latin American Cooperative Group
Hemostasis and Thrombosis (CLAHT). Bs As, 18/20-4-79.
- XXIV Meeting of the Society for Research Argentina Clinic. Mar del Plata, November 1979.
- Sixth Latin American Congress of Hemostasis and Thrombosis.
Latin American Cooperative Group of Haemostasis and Thrombosis. Buenos Aires, October 1980.
- Week Haematology Argentine-British. Hematologic Research Institute "
Mariano R. Castex", National Academy of Medicine. Buenos Aires, September 1980.
- VII Ibero-American Meeting of Haemostasis and Thrombosis, San Jose, Costa Rica, July 1981.
- VII International Congress on Thrombosis and Haemostasis. Toronto (Ontario), Canada, July 1981.
- XXVI Scientific Meeting of the Sociedad Argentina de Investigación Clínica. Mar del Plata, November 1981.
- Buenos Aires '81 Symposium. "Lipids, Platelets and Thrombosis."
Latin American Cooperative Group of Haemostasis and Thrombosis. Bs As, 16/18-11-81.
- German Week Haematology. Buenos Aires, October 1982.
- Buenos Aires '83 Symposium. "Antithrombotic Therapy." Latin American Cooperative Group
Haemostasis and Thrombosis. Buenos Aires, September 1983.
- VIII Latin American Congress of Hemostasis and Thrombosis. VIII Workshop Latin American Cooperative Group
Hemostasis and Thrombosis. Montevideo, Uruguay, in September 1983.
- XXVIII Scientific Meeting of the Sociedad Argentina de Investigación Clínica. Mar del Plata, November 1983.
- Buenos Aires '84 Symposium. "Wall Vascular-Platelet Interaction."
Latin American Cooperative Group of Haemostasis and Thrombosis. Buenos Aires, August 1984.
- XX Congress of the International Society of Hematology. Bs As, 1/7-9-84.
- Week Haematology Scandinavia. Buenos Aires, July 1985.
- Buenos Aires '85 Symposium. "Adhesiveness and platelet aggregation, and Clinical Pharmacology." Latin American Cooperative Group
Haemostasis and Thrombosis. Buenos Aires, September 1985.
- I Scientific Meeting 1986. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Fibrinolysis. Bs As, 03.29.1986.
- II Meeting Scientific, 1986. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: von Willebrand disease. Bs As, 06.26.1986.
- Buenos Aires '86 Symposium. "Thrombolytic Therapy: Tissue Plasminogen Activator." Latin American Cooperative Group
Haemostasis and Thrombosis. Buenos Aires, September 1986.
- III Scientific Meeting 1986. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Experiences and methodological applications of Group members in different
CAHT International Centers in 1986. Buenos Aires, October 1986.
- XI Congress of the Latin American Association of Pharmacology. II Congress of the American
Clinical Pharmacology and Therapeutics. Bs As, 24/29-11-86.
- I Scientific Meeting 1987. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Use of blood in patients with congenital and acquired disorders of hemostasis.
Indication and treatment regimens. Bs As, 06.05.1987.
- International Congress on Thrombosis and Haemostasis. Brussels, Belgium, 6/10-7-87.
- II Scientific Meeting 1987. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Methods of diagnosis and treatment goals in thrombosis. Bs As, 31.07.1987.
- III Scientific Meeting 1987. Group Argentine cooperative Haemostasis and Thrombosis.
Theme: Disorders of hemostasis in malignant diseases. Bs As, 11.09.1987.
- Buenos Aires '87 Symposium. "Physiology and treatment of thrombosis." Latin American Cooperative Group
Haemostasis and Thrombosis. Bs As, 14/16-9-87.
- IV Scientific Meeting 1987. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Antithrombotic therapy in various medical conditions. Cordoba, 11/27/1987.
- I International Symposium on Thrombosis. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Bs As, 11/13-4-88.
- Satellite Meeting of the First International Symposium Thrombosis. Argentinean Cooperative Group
Haemostasis and Thrombosis. Mendoza, 14/15-4-88.
- Franco-Argentine Symposium of Hematology. Bs As, 21/22-4-88.
- Medical Conference on Thrombosis and Anticoagulation. Roemmers Foundation. Bs As, 13/14-6-88.
- II Scientific Meeting 1988. Argentinean Cooperative Group of Haemostasis and Thrombosis. Topic:
Desmopressin (DDAVP) in the treatment of disorders of hemostasis in
different pathologies. Bs As, 07.01.1988.
- Buenos Aires '88 Symposium. "Endothelial cells and thrombosis."
Latin American Cooperative Group of Haemostasis and Thrombosis. Bs As, 14/16-9-88.
- III Scientific Meeting 1988. Argentinean Cooperative Group of Haemostasis and Thrombosis. Topic:
antiphospholipid Inhibitors, clinical aspects, diagnosis and treatment. Bs As, 28.11.1988.
- II Educational Symposium on Antithrombotic Prophylaxis and Therapy. II Scientific Meeting of the Argentinean Cooperative Group
Haemostasis and Thrombosis. Bs As, 4/5-4-89.
- Satellite Meeting of the Second Symposium on Prophylaxis and Therapy Education Antithrombotic; Argentinean Cooperative Group
Haemostasis and Thrombosis. Cordoba, 07/04/1989.
- International Symposium I Argentinean Cooperative Group of Thrombosis and Haemostasis and
Cooperative Group Latin American Hemostasis and Thrombosis. Argentina Society for the Study of Thrombosis
. Bs As, 29.05.1989.
- III Scientific Meeting 1989. Argentinean Cooperative Group of Haemostasis and Thrombosis.
"Causes of congenital thrombotic disease." Bs As, 04.08.1989.
- III International Symposium Argentinean Cooperative Group of Haemostasis and Thrombosis. "
Trends in treatment of thrombotic disorders." Bs As, 9/11-5-90.
- Buenos Aires '90 Symposium. "Atherogenesis and thrombogenesis, Physiology and Treatment." Latin American Cooperative Group
Haemostasis and Thrombosis. Bs As, 5/7-9-90.
- II Meeting Science 1990. Argentinean Cooperative Group of Haemostasis and Thrombosis.
"Participation of leukocytes in the regulation of hemostasis. Bs As, 10.18.1990.
- III Scientific Meeting 1990. Argentinean Cooperative Group of Haemostasis and Thrombosis.
"Modulators endothelial and thrombosis." Bs As, 12.17.1990.
- IV International Symposium Argentinean Cooperative Group Haemostasis and Thrombosis. "New therapeutic perspectives
thrombosis." Bs As, 9/11-4-91.
- II Scientific Meeting 1991. Argentinean Cooperative Group of Haemostasis and Thrombosis.
"Thrombosis and Haemostasis. Bs As, 06.14.1991.
- Conference "Thrombosis Factors and Growth and Proliferation of Smooth Muscle Cells
in atherosclerosis and coronary restenosis. "Argentinean Cooperative Group
Hemostasis and Thrombosis, Bs As, 16.09.1991.
- Symposium:" Endothelium, coagulation and thrombosis. Determinants in the development of atheromatous lesions
and restenosis. Atheroma regression. "
Argentinean Cooperative Group Haemostasis and Thrombosis, Bs As, 09.20.1991.
- Measurement of eicosanoids by EIA. Argentino Cooperative Group of Hemostasis and Thrombosis.
Bs As, 11.10.1991 .
- III Scientific Meeting 1991. Argentinean Cooperative Group Haemostasis and Thrombosis.
Topic: Thrombosis in Neurology. "Bs As, 11.06.1991.
- XII International Congress of the Latin American Cooperative Group of Haemostasis and Thrombosis.
Buenos Aires, from 18 to 11/22/1991.
- I Scientific Meeting of 1992 . Argentinean Cooperative Group Haemostasis and Thrombosis.
Theme: "Coagulation and Thrombosis in Obstetrics." Bs As, 04.10.1992.
- V International Symposium Argentinean Cooperative Group of Haemostasis and Thrombosis. "New Perspectives on Diagnosis
Therapeutic and antithrombotic. "Bs As, 6/8-5-92.
- Scientific Meeting (workshop type.) Argentinean Cooperative Group Haemostasis and Thrombosis.
Topic: Bleeding Time." Bs As, 8-5 - 92.
- II Scientific Meeting 1992. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: "Low Molecular Weight Heparin and antithrombin Therapeutics." Bs As, 06.12.1992.
- III Scientific Meeting 1992. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Theme: "Hepatic and Coagulation." Bs As, 08.14.1992.
- 56 ° Annual Scientific Triduum. Biochemistry Association Argentina. Buenos Aires, 26 to 10/31/1992.
- Symposium Buenos Aires 1992. Latin American Cooperative Group of Haemostasis and Thrombosis.
Theme: "Coronary Angioplasty." Buenos Aires, 1, 12/02/1992.
- II Scientific Meeting 1993. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Oral anticoagulation: an update. " Bs As, 18.06.1993.
- IIHema Scientific Meeting of the National Academy of Medicine and the Hemophilia Foundation of Argentina. Topic: "Present and future treatment of hemophilia." Bs As, 11.08.1993.
- Buenos Aires 1993 Symposium. CAHT Group. Theme: "heparin. thrombolysis and / or angioplasty in AMI
. " Bs As, 15 and 11/16/1993.
- II CAHT Group Scientific Meeting, jointly with the Stroke Council of the SAC, Bs As, 24.04.1995:
"Coronary artery disease in patients with TIA and stroke: prevalence, implications
prognostic and therapeutic. "
05/22/1995: Risk of stroke in cardiac lesions.
28/11/1995: Thrombophilia family, clinical, diagnostic and therapeutic approach with special
DVT, arterial thrombosis and embolism and ischemic stroke in young people.
- Conference Dr Dahlback (Sweden): Resistance to activated protein C as risk factor for venous thrombosis
. Lab Chromogenics and CAHT group, Bs As, 11.20.1995.
- Joint Scientific Meeting Saetre and Society of Internal Medicine of Buenos Aires
Tribute to the Masters of Hematology. 09/05/1996.
- Italo-Argentine Conference of Hematology, IIHema, National Academy Medicine, 3/4-10-96.
- Ia Scientific Meeting 1997. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Topic: Pathology thrombotic pregnancy. " Buenos Aires, 23/05/1997.
- IIa Scientific Meeting 1997. Argentinean Cooperative Group of Haemostasis and Thrombosis.
Theme: "prothrombotic states." Buenos Aires, 11/07/1997.
- IIIa Argentine Congress of Hemostasis and Thrombosis. CAHT Group, Buenos Aires, 18 to 20/11/1998
- XLIII Annual Meeting of the Sociedad Argentina de Investigación Clínica, Mar del Plata, 26/29/11/98.
- Ibero-American Symposium of Hematology. IIHema Academy Of Medicine, 14/16-4-99
- International Symposium on Hodgkin Lymphoma. Review and Update, Roemmers Foundation,
08/06/1999.
- Ia. CAHT Group Scientific Meeting 2000. Theme: "thrombocytopenia." 05/19/2000.
- Scientific Meeting of the Group IIa CAHT 2000. 06/30/2000.
- XLV Annual Meeting of the Sociedad Argentina de Investigación Clínica, Mar del Plata, 22-25/11/2000.
- British Days of Hematology. Total hours: 13. IIHema, National Academy of Medicine;
April 2001.
- International Symposium on plurimetabolic syndrome; Foundation Roemmers, 04/26/1901.
- International Symposium "Hemolytic Uremic Syndrome," National Academy of Medicine, June 2001.
- International Symposium on Vascular Disease TTP; Roemmers Foundation, 1/10/1919.
- XLVII Annual Meeting of the Sociedad Argentina de Investigación Clínica, Mar del Plata, 22-23/11/2002.
- Symposium on Hemostasis and Thrombosis, CAHT Group, Margulies Foundation, 29/11/1902.
- Attended and presented papers at the XIX Congress and 49th Annual SSC Meeting of the International Society on Thrombosis
and Haemostasis. Birmingham, UK, 12-18 July 2003.
- Symposium Hemostasis and Thrombosis, CAHT Group, Margulies Foundation, 08/08/1903.
- Joint Congress of Biomedical Societies, Mar del Plata, 16-20/11/04.
- Fourth Scientific Meeting CAHT Group, Del Viso, Pilar, 11/26/1904.
- North American Conference of Hematology. IIHema, National Academy of Medicine, 14 and 04/15/1905.
- International Symposium on Hemorrhagic and Thromboembolic Complications in Oncology.
Roemmers Foundation, National Academy of Medicine, 07/01/1905.
- International Symposium on glycoproteins. Tribute to Luis F. Nobel Prize Leloir. National Academy of Medicine and National Academy of Sciences, Physical and Natural Sciences. 09/12/1906.
- Molecular Immunology of Protozoan Infections. HHMI, EMBO, WHO, ANPCyT, CONICET, UNU-
BIOLAC, CABBIO, 19 and 03/20/1907.
- Presentation Dr. Jorge di Paola (University of Iowa) 03/29/1907, Lanari Institute.
Topic: genetic modifiers von Willebrand factor
- Canadian Conference of Hematology. IIHema. National Academy of Medicine, 19 and 20/04/1907.
- XX Congress of the Latin American Group of Haemostasis and Thrombosis (CLAHT) -
Group VIII Argentine Congress of Hemostasis and Thrombosis (CAHT). From 23 to April 26, 2008. Graduate courses
- Course Thrombosis Research Institute Hematologic "Mariano R. Castex", National Academy of Medicine. Buenos Aires. 12/22-05-78.
- Course Haemostasis, Hematological Research Institute 'Mariano R. Castex ", National Academy of Medicine. Buenos Aires. 24/28-5-78.
- Theoretical and Practical Course "Research Methodology and Biostatistics, IIHema, National Academy of Medicine, on days 1, 8, 15, 22 and 29/09/1999.
- Course organized by PE Biosystems of Brazil "DNA Sequencing on the ABI PRISM 310 Genetic Analyzer."
8/12-11-1999
- Theoretical and Practical Course "Introduction to flow cytometry" total hours: 8. IIHema, National Academy of Medicine, 7 and 8 June 2001.
- Basic Biostatistics course of 80 class hours, with a final exam passed. Epidemiologic Research Center, National Academy of Medicine, July 2001.
- Advanced Biostatistics course of 70 class hours, with a final exam passed.
Epidemiologic Research Center, National Academy of Medicine, November 2001.
- Molecular Diagnostics in medicine. Its application to genetic diseases, cancer and infectious
, with a final exam passed. Centre for Research on Porphyrins and Porphyrias
(CIPYP). Department Biological Chemistry. FCEyN, UBA 9/17-12-02.
- Meta-analysis. Epidemiologic Research Center, National Academy of Medicine, the
16 / 3 18 / 5 2004.
- Principles and applications of RNA interference (siRNA), Sociedad Argentina
Research Clinical Genetics Council, 08/31/2005.
- Molecular Diagnostics in medicine. Its application to genetic diseases, cancer and infectious 2005 with approved final assessment (90 hours duration). Centre for Research on Porphyrins and Porphyrias (CIPYP). Department of Biological Chemistry. FCEyN, UBA. 8/25-11-05.
- Minicourse: Biosynthesis Combinatorics: New Technology for Generation of Therapeutic Agents. Carried out during the 50th Annual Meeting of the Sociedad Argentina de Investigación Clínica, Mar del Plata, 29 to December 2, 2005.
- Mini-Course: Analysis and Interpretation of Microarray Data. GE Healthcare Bio-Sciences. Molecular Classification of Prostate Tumors Using Intronic Arrays. Carried out during the 50th Annual Meeting of the Sociedad Argentina de Investigación Clínica, Mar del Plata, 29 to December 2, 2005.
- Minicourse: Mutagenesis Techniques Applied to the Study of Protein Structure and Function. Carried out during the 50th Annual Meeting Sociedad Argentina de Investigación Clínica, Mar del Plata, 29 to December 2, 2005.
PUBLICATIONS in refereed journals
1 - Meschengieser S, Woods AI, Schattner M, Lazzari MA: Association of hemorrhagic and thrombotic phenomena in myeloproliferative syndromes. Medicina (Buenos Aires) 41:267-273, 1981.
2 - Woods AI, Sutton NM, Kempf A, Casillas G, Lazzari MA: Use of plasma ultrasonicated as diluent in the art metering ristocetin cofactor. Blood 30:999-1006, 1981.
3 - Woods AI, Vargas J, Berri G, Kreutzer G, Meschengieser S, Lazzari MA: Antithrombotic therapy in Children and adolescent. Thromb. Res. 42:289-301, 1986.
4- Woods AI and Lazzari MA: Aspirin effect on platelet antiplasmins release. Thromb Res. 47:269-277, 1987.
5- Meschengieser S, Blanco A, Woods AI, Maugeri N, Fernández J, Dupont J, Lazzari MA: Intraplatelet levels of FvW:Ag and fibrinogen in myeloproliferative disorders. Thromb. Res. 48:311-320, 1987.
6- Woods AI. and Lazzari MA: Aspirin effect on the release of plasminogen activators inhibitors by human platelets. Thromb. Res. 52:119-125, 1988.
7- Farías C, Kempfer A, Blanco A, Woods AI, Lazzari MA: Visualization of the multimeric structure of von Willebrand Factor by immunoenzymatic stain using Avidin-Peroxidase Complex instead of Avidin-Biotin-Peroxidase Complex. Thromb. Res. 53:513-518, 1989.
8- Woods AI, Frontroth JP, Lazzari MA: Effect of protease inhibitors on the ristocetin cofactor activity. Rev. Iberoamer. Tromb. Hemostasia. 2:47-48, 1989.
9- Woods AI, Nadra C, Lazzari MA: The effect of venous occlusion on the PAI and anitplasmins release from platelets. Thromb. Res. 56:491-495, 1989.
10- Grand B, Blanco A, Riveros D, Ventura A, Maugeri N, Woods A, Lazzari MA: Congenital Factor XIII deficiency associated with von Willebrand's disease. Am. J. Hematol. 35:208-209, 1990.
11- The Ridogrel Versus Aspirin Patency Trial (RAPT). Randomized Trial of Ridogrel, a combined TXA2 Synthase Inhibitor and TXA2/ prostaglandin endoperoxide receptor antagonist, versus aspirin as adjunct to thrombolysis in patients with acute myocardial infarction. Circulation 89:588-595, 1994.
12- Woods AI, Santarelli M, Lazzari MA: The inhibitory effect of platelets on fibrinolysis in diabetic patients. Thrombosis Research 76:391-396,1994.
13- Farias CE, Gimenez JC, Kempfer AC, Singer E, Woods AI, Santarelli MT, Lazzari MA. Ionizing radiation increases concentration of plasma von willebrand factor in cebus apella paraguayanus monkeys. Thrombosis Research 88,81-87,1997.
14- Kempfer AC, Silaf MR, Farias CE, Carballo GA, Woods AI, Lazzari MA. Binding of von Willebrand factor to collagen by flow cytometry. Am. J. Clin Pathol. 111,418-423,1999.
15- Woods AI, Meschengieser SS, Blanco AN, Salviú MJ, Farias CE, Kempfer AC, Lazzari MA. Clinical features and laboratory patterns in a cohort of consecutive Argentinian patients with von Willebrand`s disease. Haematologica 86,420-427;2001.
16- Sánchez-Luceros A, Meschengieser SS, Woods AI, Blanco AN, Kempfer AC, Casais P, Salviú MJ,
Lazzari MA. Acquired von Willebrand factor abnormalities in myeloproliferative disorders and other
hematologic diseases: a retrospective analysis by a single institution. Haematologica 87:264-70; 2002.
17- Sanchez-Luceros A, Meschengieser SS, Votta R, Casais P, Woods AI, Nadal MV, Salviú MJ, Lazzari MA. Factor VIII and von Willebrand factor changes during normal pregnancy and puerperium. Blood Coagulation and Fibrinolysis 14:647-651; 2003.
18- Kempfer AC, Farias CE, Amaral MM, Silaf MR, Woods AI, Carballo GA, Lazzari MA. Control of von Willebrand factor multimer size by a fibronectin related substance. Blood Coagulation and Fibrinolysis 14:441-8; 2003.
19- Sanchez-Luceros A, Farías CE, Amaral MM, Kempfer AC, Votta R, Marchese C, Salviú MJ, Woods AI, Meschengieser SS, Lazzari MA. Von Willebrand factor-cleaving protease (ADAMTS 13) activity in normal non-pregnant women, pregnant and delivery women. Throm Haemost 2004;92:1320-6.
20- Favaloro EJ, Lilicrap D, Lazzari MA, Cattaneo M, Mazurier C, Woods A, Meschengieser S, White A, Kempf AC, Hubbard A, Chang A. von Willebrand Disease: Laboratory aspect of diagnosis and Treatment. Haemophilia 2004; 10: (suppl.4), 164-168.
21 - Casais P, Carvalho GA, Woods AI, Kempf AC, Farias CE Grosso SH, Lazzari MA. Within sustitution encoded R924Q Exon 21 of the von Willebrand factor gene related to wild Bleeding phenotype. Thromb Haemost 2006; 96:228-30.
22 - Sánchez-Luceros A; Meschengieser SS; turds K, Ariza A, Woods AI, couples P, White NA, AC Kempf, Maria A Lazzari. Clinical Evaluation of the safety of desmopressin in women During Pregnancy With A low plasma von Willebrand factor level and Bleeding history. Thromb Res 2007; 120.387-90.
PUBLICATIONS Refereed Journal without
1 - Woods AI, Meschengieser S, Lazzari MA: Platelet function in diabetic patients. Bulletin of the National Academy of Medicine. 59:297-302, 1981.
2 - Meschengieser S, Woods AI, Lazzari MA: Oral anticoagulation in pregnant patients with mechanical valve prostheses. Bulletin of the National Academy of Medicine. 65:359-363, 1987.
3 - Meschengieser S, Bermejo E, Maugeri N, Woods AI, Blanco A, Pieroni G, Lazzari MA: Use of DDAVP in patients with von Willebrand disease and thrombocytopathies. Bulletin of the National Academy of Medicine. 67:135-145, 1989.
4 - Bermejo E, Meschengieser S, Woods A, Peñalba L, Pieroni G, Lazzari MA: DDAVP: Changes of hemostasis caused by its use. Prensa Médica Argentina 79:376-380,1992.
5 - Farias CE, Kempf AC, Woods A, Lazzari MA. Multimeric population analysis of platelet von Willebrand factor after DDAVP administration. Bulletin of the National Academy of Medicine 71:509-515, 1993.
6 - Peñalva L, Dib Ashub M, Blanco A, Woods AI, Lazzari MA. Fibrinolytic response to 1 - deamino-d-arginine vasopressin (DDAVP) in patients with impaired hemostasis. Bulletin of the National Academy of Medicine 71:81-84, 1993.
7 - Woods A, Meschengieser S, White, AN, Salviú MJ, Farias CE, Kempf BC Lazzari MA. Von Willebrand disease, the most common bleeding disorder. Bulletin of the National Academy of Medicine 79:275-290, 2001.
ABSTRACTS PUBLISHED IN JOURNALS OF CONFERENCE NATIONAL PERIODICAL
1 - Lazzari MA, Schattner M, Woods AI, Arce R: Controls of antithrombotic therapy. Medicina (Buenos Aires) 39 (6): 715, 1979 (to 15).
2 - Meschengieser SS, Woods AI, Lazzari MA: Platelet function in diabetic patients. Medicina (Buenos Aires) 41 (6): 692, 1981 (to 88).
3 - Woods AI, Lazzari MA: fibrinolytic activity in diabetic patients. Medicina (BsAs) 41 (6): 693, 1981 (to 89).
4 - Woods AI, Sutton N, Lazzari MA: Use of plasma ultrasonicated in assessing the ristocetin cofactor. Medicine (Buenos Aires) 43 (6 / 2): 828, 1983 (to 205).
5 - White A, Meschengieser SS, Woods AI, Farias C, Maugeri N, Bermejo E, Pieroni G, Ventura A, Lazzari MA: myeloproliferative syndromes (SMP) and acquired von Willebrand (vWA). Medicine (Bs.As) 48 (6): 624, 1988 (a.192)
6 - Kampfer A, Frontroth J, Farias C, Bermejo E, Woods AI, Lazzari MA: Involvement of serum components in the adhesion of von Willebrand factor (vWF) to the subendothelium in perfusion chamber. Medicina (Buenos Aires) 49 (6): 438, 1989 (a.109)
7 - White A, Grand B, Pieroni G, Peña L, Voto L, Ventura A, Woods A, Lazzari MA: Complex factor VIII / von Willebrand factor (VIII / vW) in pregnancy-induced hypertension. Medicine 52 (sup II): 391, 1992.
8 - Kampfer A, Farias C, Woods A, Frontroth JP, Bermejo E, Lazzari MA: multimeric structure of von Willebrand factor (vWF), platelet after administration of DDAVP. Medicine 52 (suppl II): 392, 1992.
9 - Kampfer A, Farias C, Woods A, Frontroth JP, Bermejo E, Lazzari MA: multimeric structure of von Willebrand factor (vWF) under high shear rate. Medicine 52 (suppl II): 391.1992.
10 - Woods AI, Kampfer AC, Effect of collagen and aspirin on the release of PAI-1 and intraplatelet t-PA. Medicine 53 (suppl II), 43.1993 (a 95).
11 - Bermejo E, Woods AI, Meschengieser S, Lazzari MA. Prolongation of bleeding time and platelet function abnormalities in patients with von Willebrand disease. Medicine 53 (suppl II); 74, 1993 (to 155).
12 - Szwarcer E, Giuliani AR, White A, Palumbo G, Bartomioli M, Orlando S, Woods A, Aguero G, Baitch L, Villafañe S, Lanari E, Amaya ED, Diaz R, ECLA Study Group 2, Bs As Argentina. Relationship between parameters of haemostasis, coronary patency and bleeding in the 2-RAPT ECLA study. 1 Argentine Congress Hemostasis and Thrombosis, group CAHT, Mar del Plata, 7 to 10/09/1994.
13 - Farias CE, Woods AI, Bermejo E, Meschengieser S, Kampfer AC, Lazzari MA. Patients with von Willebrand disease type 1. Expression of different subtypes according to the intraplatelet von Willebrand factor. Medicine 54, 494, 1994 (to 31).
14 - Farias CE, Kempf AC, Woods AI, Bermejo E, Lazzari MA. Comparisons of the Platelet fibronectin in patients with von Willebrand disease type 1. normal platelet and platelet discordant, before and after infusion of desmopressin. Medicine 54, 494, 1994 (to 32).
15 - Farias CE, Kempf AC, Woods AI, Lazzari MA. Effect of blood group and sex in the content of von Willebrand factor (vWF) intraplatelet. Medicine 55, 83, 1995.
16 - Farias CE, AC Kampfer, Meschengieser S, Woods AI, Blanco A, Lazzari MA. Monitoring the response to desmopressin (DDAVP) in patients with von Willebrand (vW) using the collagen binding assay. Medicine 56, 621.1996 (A261)
17 - Kampfer AC, Farias CE, MR Silaf, Carballo GA, Woods AI, Lazzari MA. Detection of the binding of von Willebrand Factor (vWF) to collagen and vWf: Ag by flow cytometry. Medicine 57 (suppl 4): 2.1997 (8).
18 - Farias CE, AC Kampfer, Meschengieser S, Carballo GA, Silaf MR, Blanco A, Woods AI, Lazzari MA. Usefulness of a functional assay based on the binding of von Willebrand factor (vWf) to FVIII to classify patients with von Willebrand disease. Medicine 57 (suppl 4): 4.1997 (at 14).
19 - Woods AI, Maugeri N, Kampfer AC, Lazzari MA. Effect of low shear rate on the release of PAI-1 Platelet; their inhibition with aspirin. Medicine 57 (suppl 4): 5.1997 (at 17).
20 - Woods AI, Blanco, AC Kampfer, Meschengieser SS, Lazzari MA. Our experience in the diagnosis of von Willebrand disease. Medicine 58 (5 / 2), 668.1998 (a 460)
21 - Woods AI, Blanco, AC Kampfer, Meschengieser SS, Lazzari MA.Penetrancia the phenotype of the disease von Willebrand type 1 and 2A. Medicine 58 (5 / 2): 668, 1998 (to 461)
22 - Farias CE, Kempf AC, Carballo GA, Silaf MR, Casais P, Woods A, Grosso S, Lazzari MA. Frequency of the Arg 91 Gln mutation in von Willebrand disease type 2N. Medicine 58 (5 / 2): 668, 1998 (to 462)
23 - Sanchez Luceros A, Ariza A, Casais P, Woods AI, Meschengieser SS, Lazzari MA. Use of desmopressin in pregnant women with von Willebrand disease. Medicine 59 (5 / 2): 560, 1999 (to 9).
24 - Woods AI, Kempf A, Silaf M, Farias C, Grosso S, Carballo G, Lazzari M. Inhibition of ristocetin cofactor activity by the presence of EDTA. Medicine 60 (5 / 2): 790; 2000 (241).
25 - Farias C, Kempf A, Sanchez Luceros A Silaf M, Woods A, Carvalho G, Lazzari M. Extra-large multimers of von Willebrand factor in normal pregnant women in the third quarter. Medicine, 60 (5 / 2): 790, 2000 (242).
26 - Kempf AC, Carvalho GA, Farias C, Silaf MR, Amaral MM, Woods AI, Lazzari MA. Relationship between fragments of fibronectin (Fn) high affinity for heparin and the protease that the Clive
von Willebrand factor (VWFCP). Medicine 61 (5 / 2): 770, 2001 (the 393).
27 - Kempf AC Farias C, Silaf MR, Carvalho GA, Amaral MM, Woods AI, Lazzari MA. Differentiation between the protease related to fibronectin (PRFN) described other proteases that cleave the von Willebrand factor (VWF) and matrix metalloproteinases (MMP). Medicine 62 (5): 409, 2002 (to 65).
28 - Farias C, Kempf A, Amaral MM, Woods AI, Nadal V, Carballo G, Meschengieser S, Lazzari MA. Virtual decrease of protease cleaving von Willebrand factor (VWFCP) after administration of desmopressin (DDAVP). Medicine 62 (5): 442, 2002 (to 212).
29 - Carballo GA, Woods AI, Farias C, Kampfer AC, Lazzari MA. Von Willebrand disease type 2N. Molecular biology. Medicine 62 (5): 442, 2002 (to 213).
30 - Woods AI, Meschengieser SS, White A, Lazzari MA. Risk of major bleeding in patients with von Willebrand disease type 1. Medicine 62 (5): 444, 2002 (to 221).
31 - Woods AI, Nadal MV, Meschengieser SS, Blanco AN, Lazzari MA. Influence of intra-platelet VWF in the presence of major bleeding in surgery. Medicine 63: 654, 2003 (to 526).
32 - Kampfer AC, Farias C, Sanchez-LUCERO A, Amaral MM, Grosso S, Carballo G, Woods A, Lazzari MA. Discordance between levels of von Willebrand factor (VWF) and factor VIII (FVIII) hemophilia or not related to the variant 2 N von Willebrand disease. Medicine 63:654, 2003 (to 528).
33 - Nadal MV, Blanco AN, Woods AI, Meschengieser SS, Grosso SH, Salviú MJ, Lazzari MA. Von antigen Platelet Willebrand (VWF: Ag-i) in patients with mucocutaneous bleeding and normal levels of plasma von Willebrand factor (VWF: Ag). Medicine 63: 654, 2003 (to 529).
34 - Carballo GA, Woods AI, Kampfer AC, Farías CE, Amaral MM, Lazzari MA. R924Q substitution in a family with a slight decrease of FVIII binding to VWF. Medicine 63:656, 2003 (to 534).
35 - Carballo GA, Woods AI, Vera E, Casais P, Aletti G, Farías CE, Kempf AC, Lazzari MA. Genetic alterations of von Willebrand factor in 22 family members in Argentina. Medicine 64: (suppl II), 2004 (to 400).
36 - Woods AI, Blanco AN, Nadal MV, Meschengieser SS, Grosso SH, Lazzari MA. Fibrinolytic response to DDAVP in patients with type 1 VWD likely without major bleeding in surgery. Medicine 64: (suppl II), 2004 (to 678).
37 - Keller L, Woods AI, Kampfer AC, Farías CE, Grosso N, Lazzari MA. Validation of the Conformation Sensitive Gel Electrophoresis (CSGE) as a method of screening of exon 22 of von Willebrand factor in the diagnosis of von Willebrand disease (VW) 2N. Medicine 65: (suppl II), 2005 (to 65).
38 - Marti A, Casais P, Kampfer AC, Woods A, White A, Meschengieser S, Lazzari MA. Von Willebrand disease subtype 2B and management of labor. XVII Argentine Congress of Hematology. 60 th Anniversary of the Society Argentina Hematology. Córdoba. 2-5.11.2005.
39 - Keller L, Woods AI, Farías CE, Sanchez-LUCERO A, Grosso N, Kampfer AC, Powazniak YP, Calderazzo JC, Lazzari MA. Association of von Willebrand type 2N and type 2M Vicenza. Medicine 66: (supl.II), 2006 (to 162).
40 - Carrivale M, Kampfer AC, Woods AI, Vizcaychipi K, Astudillo CG, Sanchez Luceros A, Lazzari MA. Identification of patients with shortened survival of von Willebrand factor. Medicine 67 (suppl III), 2007 (to 14).
41 - Woods AI, Blanco AN, Grosso SH, JC Calderazzo, Meschengieser SS, Lazzari MA. Differential diagnosis between type 3 VWD and severe Type 1 by the response to DDAVP ELT time. Medicine 67 (suppl III), 2007 (to 9).
42 - Keller L, Kampfer AC, Woods AI, Sanchez Luceros A, Powazniak And Carrivale M, Lazzari MA. New polymorphisms in the ADAMTS gene intron 13. Medicine 67 (suppl III), 2007 (to 23).
ABSTRACTS PUBLISHED IN JOURNALS OF INTERNATIONAL CONFERENCE PERIODICAL
1 - Meschengieser S, Woods AI, Lazzari MA: Association of hemorrhagic and thrombotic phenomena in myeloproliferative syndromes. VI Congress of Hemostasis and Thrombosis Latinomaricano. Bs.As, October 1980. Abstract book p.13.
2 - Woods AI, Meschengieser S, Sutton M, Lazzari MA: Correlation between ristocetin cofactor levels and various parameters of platelet aggregation in diabetic patients. VI Latin American Congress of Hemostasis and Thrombosis. Buenos Aires, October 1980. Abstract book p.12.
3 - VIII International Congress of Haematology. Montreal, Canada, October 1980. Abstract Book p. 145.
4 - Woods AI, Lazzari MA: Platelets and fibrinolysis. VII Latin American Meeting of Haemostasis and Thrombosis. San José, Costa Rica, July 1981. Book of Abstracts.
5 - Woods AI, Meschengieser S, Sutton M, Lazzari MA. Platelet function parameters in diabetic Patients. Thromb Haemost 1981, 46 (1). (To 1413).
6 - Meschengieser S, Woods AI, Schattner M, Lazzari MA: Antiplatelet action of ticlopidine on thrombotic Normal Subjects and Patient. VII International Congress on Thrombosis. Haemostasis 1982, 12:159.
7 - Lazzari MA, Woods AI, Sutton N: Use of ultrasonicated for the dosage of plasma ristocetin cofactor. IX International Congress on Thrombosis and Haemostasis. Stockholm, Sweden. 3 / 8 Julio 1983. Thromb. Haemost 1983; 50 (1): 224 (the 764).
8 - Woods AI, Vargas J, Berri G, Kreutzer G, Meschengieser S, Lazzari MA: Our experience in the management of Anticoagulation in children and Adolescent. XX Congress of International Society of Hematology. Bs.As., September 1984. Abstracts Book.
9 - Woods AI, Lazzari MA: Action of aspirin on platelet antiplasmin release. XI Congress of the Association of Pharmacology and II Latinomamericana Congress of the American Society for Clinical Pharmacology and Therapeutics. Buenos Aires, 24/29 November 1986. Book of Abstracts (a.227).
10 - Meschengieser S, Woods AI, Lazzari MA: Anticoagulation in pregnancy in Patients with cardiac valve prosthesis. Thromb. Haemost. 1987, 58 (1): 130 (to 467).
11 - Woods AI, Lazzari MA: Aspirin effect on platelet antiplasmins release. Thromb. Haemostas.1987, 58 (1): 189 (at 698).
12 - Woods AI, Lazzari MA: Aspirin failure to Inhibit the release of plasminogen Activator inhibitors by human Platelets. Thromb. Haemost. 1987, 58 (1): 94 (at 327).
13 - Lazzari MA, Woods AI: Platelets and fibrinolysis. Conference. X Congress of Hemostasis and Thrombosis. Latin American Cooperative Group of Haemostasis and Thrombosis. Venezuela, in September 1987.
14 - Grand B, Riveros D, Blanco A, Maugeri N, Ventura A, Woods AI, Lazzari MA: Congenital factor XIII deficiency associated with von Willebrand disease. XVIII International Congress of the World Federation of Hemophilia. Madrid, Spain. 31/05/1988. Abstracts Book (50).
15 - Woods AI, Lazzari MA: Platelet antiplasmins release and calcium blockers. X International Congress on Thrombosis. Athens, Greece. 22/27-05-88. Abstract Book (200).
16- XI Congreso Latinoamericano de Hemostasia y Trombosis 1988. Grupo Cooperativo Latinoamericano de Hemostasia y Trombosis. La Habana, Cuba, 16/19-05-88. Libro de Resúmenes.
17- Woods AI, Nadra C, Lazzari MA: Platelet PAI-1 in venous stasis. VI Meeting of the Danubian League against Thrombosis and Haemorrhagic disorders. Viena, Austria, 31-05/03-06-89. Abstract Book.
18- Woods AI, Nadra C, Lazzari MA: Platelet antiplasmins in venous occlusion. VI Meeting of the Danubian League against Thrombosis and Haemorrhagic disorders. Viena, Austria, 31-5/3-6-89. Abstract Book.
19- Woods AI. Lazzari MA: Platelet PAI-1 released calcium blockers. Thromb. Haemostas. 1989; 62 (1):402 (a 596 1279).
20 - White A, Meschengieser S, Woods A, Farias C, Maugeri N, Bermejo E, Pieroni G, Ventura A, Lazzari MA: von Willebrand abnormalitites in myeloproliferative syndromes. Thromb. Haemost. 1989, 62 (1): 223 (at 690).
21 - Fondevila C, Meschengieser S, Bermejo E, Woods AI, Pieroni G, Lazzari MA. Hemostatic complications during childbirth in patients with von Willebrand disease. XII International Congress of Hemostasis and Thrombosis, Buenos Aires, November 1991. Book of Abstracts. (To 112).
22 - Fondevila C, Meschengieser S, Bermejo E, Woods AI, Pieroni G, Lazzari MA. Platelet adhesiveness in von Willebrand disease during pregnancy. XII International Congress Hemostasis and Thrombosis, Buenos Aires, November 1991, Book of Abstracts. (To 111).
23 - Woods AI, Lazzari MA antifibrinolytic platelet activity in diabetic patients. XII International Congress of Hemostasis and Thrombosis. Buenos Aires, November 1991. Book of Abstracts (to 155).
24 - Szwarcer E, Giuliani AR, White A, Palumbo G, Bartomioli M, Orlando S, Woods A, Aguero G, Baitch L, Villafañe S, Lanari E, Amaya ED, Diaz R, ECLA 2, Study Group: Relationship of Hemostasis Parameters to coronary patency and bleeding in 2 RAPT ECLA Trial XVth Congress of the European Society of Cardiology, Nice, France, August 29-September 2, 1993.
25 - Woods AI, Kampfer AC, Lazzari MA. Effect of collagen and aspirin on the intraplatelet PAI-1 and t-PA release. XXV Congress of the I.S.H. Cancun, Mexico. Abril 17/22-1994 (a 604).
26- Meschengieser S, Farias CE, Woods AI, Bermejo E, Kempfer AC, Lazzari MA. Intraplatelet von Willebrand factor and response to DDAVP. XXV Congress of the I.S.H. Cancun, Mexico abril 17/22-1994 (a 568).
27- Meschengieser S, Woods AI, Bermejo E, Lazzari MA. Different patterns in the response to intravenous DDAVP. XXV Congress of the I.S.H. Cancun, Mexico. abril17/22-1994 (a 566).
28- Woods AI, Schattner M, Lazzari MA. PAI-1 and antifibrinolytic activity released by platelets treated with cationized human IgGs. Thromb. Haemostas. 1995; 73 (6). (a 419).
29- Farias C, Meschengieser S, Woods AI, Bermejo E, Funes S, Kempfer AC, and Lazzari MA. Relationship between intraplatelet von Willebrand factor and response to DDAVP. XV Congress of the ISTH. Jerusalem, Israel, junio 11-16, Thromb. Haemostas. 1995; 73 (6). (a 1002).
30- Kempfer AC, Farias CE, Woods AI, Cardoso MA, Lazzari MA. The effect of high shear rat and a plasma fractyion on von Willebrand factor multimers size. Thromb. Haemostas. 1995; 73 (6). (a 1013).
31- Woods AI, Maugeri N, Kempfer AC, Lazzari MA. Effect on the low shear rate on the release of intraplatelet PAI-1. Its inhibition with aspirin. Thromb. Haemostas. 1997; 77 (suppl). (a PS-469)
32- Farias CE, Kempfer AC, Carballo GA, Blanco A, Woods AI, Meschengieser S, Lazzari MA. Laboratory monitoring of Desmopressin (DDAVP) using collagen binding assay (CBA). Thrombo. Haemostas. 1997, 77 (suppl) (a PS-2846).
33 - Farias CE., Kempfer AC., Carballo GA., Silaf MR., Salviu J., Woods AI., Meschengieser S., Lazzari MA. Caracterización parcial type 2N en la enfermedad de von Willebrand using fenotípicas y Técnicas genotípicas. Symposium Internacional. grupo CLAHT, Rio de Janeiro, Brasil, 16 y 07/17/1998.
34 - AC Kempfer., Silaf MR., Farias CE., Carballo GA., Woods AI., Lazzari MA. Binding of von Willebrand factor to collagen by flow citometry. Citometry Suppl. 9, 133, 1998 (in CT79).
35- Woods AI, Meschengieser S, Blanco A, Kempfer C, Santarelli MT, Lazzari MA. Our experience in the diagnosis of von Willebrand disease. XVII Congress of the International Society on Thrombosis and Haemostasis. Washington, 14-21/8/1999. Thromb Haemost (Suppl):132, 1999 (a 408).
36- Woods AI, Meschengieser S, Blanco A, Kempfer A, Lazzari MA. Prevalence of phenotype of von Willebrand disease type 1 and 2A. XVII Congress of the International Society on Thrombosis and Haemostasis. Washington, 14-21/8/1999. Thromb Haemost (Suppl):132, 1999 (a 409)
37- Woods AI, Kempfer CA, Silaf MR, Farias CE, Grosso S, Lazzari MA. EDTA-dependent low ristocetin cofactor activity. Thromb Haemost. 2001.
38 - Farías C., Kempf A., Sánchez Luceros A., Silaf M., Woods AI, Carbone G., Lazzari MA. Ultra large (UL) von Willebrand Factor multimers in Normal Pregnant Women at the third trimester. Thromb Haemost 2001 (to 785)
39 - Woods AI, Meschengieser SS, Blanco AN, Lazzari MA. Platelet VWF as a marker predictive of major bleeding in surgery. XVIII International Congress of Hemostasis and Thrombosis CLAHT Group, May 18-21th 2003, Rio de Janeiro, Brazil. (30).
40 - Grosso S, White A, Casais P, Nadal MV, Woods A, Lazzari MA. Deficiency of factor VII (FVII): frequency, clinical features and association with abnormal von Willebrand factor-VIII. XVIII Congress Internacional de Hemostasis y Trombosis Grupo CLAHT, mayo 18-21de 2003, Rio de Janeiro, Brasil. Revista Brasileira de Hematologia e hemoterapia: Vol 25 (supl1), abs. 035, pag. 33; mayo 2003.
41- Woods AI, Meschengieser SS, Blanco AN, Lazzari MA. Incidence of major haemorrhages related to surgery in type 1 von Willebrand's disease. J Thromb Haemost, 2003 (a 1682).
42- Carballo GA, Kempfer AC, Woods AI, Farias CE, Lazzari MA. Comparative study of conformation sensitive gel electrophoresis and DNA sequencing for polymorphisms and mutation detection in von Willebrand disease type 2N. J Thromb Haemost, 2003 (a 1681).
43- Farias C, Kempfer AC, Amaral MM, Woods AI, Nadal MV, Carballo GA, Meschengieser SS, Lazzari MA. Virtual decrease of von Willebrand Factor-cleaving protease (ADAMTS 13) after infusion of desmopressin (DDAVP). J Thromb Haemost, 2003 (a 1675).
44- Carballo GA, Woods AI, Vera E, Casais P, Aletti G, Farías C, Kempfer AC, Lazzari M A. Genetic Study Of von Willebrand Factor In Twenty-two Members Of An Argentinian Family. J Thromb Haemost, 2005 (a 863).
45- Woods AI; Blanco AN; Nadal MV; Meschengieser SS; Grosso SH; Lazzari MA. Evaluation of Fibrinolytic Activity and major hemorrhages in Probable Type 1 von Willebrand Disease. J Thromb Haemost, 2005 (a 263).
46- Woods AI, Keller L, Kempfer AC, Farias CR, Casais P, Lazzari MA. CSGE to scan the exon 22 of von Willebrand factor gene for diagnosising von Willebrand disease 2N. Pathophysiol Haemost Thromb 35 (1-2), Proceedings and Abstracts from the 19th International Congress on Thrombosis. Tel Aviv, Israel, May 2006.
47- Casais P, Meschengieser S, Kempfer C, Woods A, Sanchel Luceros A, Lazzari MA. Pregnancy of type 2B vWD. World Federation of Haemophilia Congress 2006. Vancouver, Canada. 30 PO 906.
48- Woods AI, Sanchez-Luceros A, Blanco AN, Grosso SH, Kempfer AC, Lazzari MA. Is there any correlation between fibrinolytic activity and bleeding score in type 1 and possible type 1 von Willebrand disease?. J Thromb Haemost 2007; vol 5, Suppl 2. (P- W 178).
49- Kempfer AC, Farias C, Powazniak Y, Woods AI, Calderazzo J, Keller L, Lazzari MA. VWF-cleaving serine proteases isolation different from ADAMTS 13. J Thromb Haemost 2007; vol 5, Suppl 2. (P- M 298).
50- Keller L, Woods AI, Kempfer AC, Farías CE, Sanchez-Luceros A, Grosso SH, Lazzari MA.
An association of Vicenza and Normandy variants von Willebrand disease (VWD). J Thromb Haemost 2007; vol 5, Suppl 2. (P- W 161).
51- Kempfer AC, Keller L, Farias C, Woods AI, Sanchez-Luceros A, Powazniak Y, Carrivale M, Lazzari MA. abstract N° 1321 - Three new polymorphisms located in the intron 2 of the ADAMTS 13 gene. J Thromb Haemost 2007; vol 5, Suppl 2 (P- M 297).
52- Sanchez-Luceros A, Meschengieser SS, Turdó K, Woods AI, Kampfer AC, Blanco AN, Lazzari MA. Clinical and biological response to desmopressin in Children with von Willebrand disease. J Thromb Haemost 2007, vol 5, Suppl 2. (P-S 248).
53 - Woods AI, Blanco AN, Grosso SH, JC Calderazzo, Meschengieser SS, Lazzari MA. Differential diagnosis between type 3 VWD and severe Type 1 by the time response to DDAVP ELT. XX Congress CLAHT Group, Eighth Congress of CAHT Group (P-135).
54 - Carrivale M, Kampfer AC, Woods AI, Vizcaychipi K, Astudillo OG, Sánchez Luceros A, Lazzari MA. Identification of patients with shortened survival of von Willebrand factor (VWF). XX Congress CLAHT Group, Eighth Congress of CAHT Group (P- 116).
55 - Gross S, Bastos L, Ingratti M, Vizcaychipi K, Woods AI, Meschengieser SS, Blanco AN, Lazzari MA. VWD and lupus anticoagulant: FBI dilution curves in the evaluation of response to DDAVP. XX Congress CLAHT Group, Eighth Congress of CAHT Group (P-084). TEACHING
- Refresher Course conducted by the National Institute of Cardiology. Buenos Aires, October 1980.
Subject: Participation of platelets in the clotting mechanism. Platelet coagulant activity.
- parallel course to the VI Congress of Hemostasis and Thrombosis. Latin American Cooperative Group of Haemostasis and Thrombosis. Buenos Aires, October 1980.
Subject: Participation of platelets in the clotting mechanism. Platelet coagulant activity.
- Refresher Course for Cardiologists, conducted by the Cardiology Division, Hospital Durand. Buenos Aires, April 1981.
Subject: Laboratory controls during treatment with oral anticoagulants.
- Haemostasis and Thrombosis course for graduates, conducted by the Association Argentina Biochemistry. Buenos Aires, October 1981.
Topics: platelet ultrastructure. Participation of platelets in the clotting mechanism. Platelet coagulant activity.
- Haemostasis and Thrombosis Course. ESSEX laboratories. Buenos Aires, October 1981.
Topic: platelet ultrastructure. Participation of platelets in the clotting mechanism.
- Haemostasis and Thrombosis course, parallel to the V Argentine Congress of Biochemistry, Biochemistry Association by Argentina. Buenos Aires, 01/06-11-81.
Topic: platelet ultrastructure. Participation of platelets in the clotting mechanism. Platelet coagulant activity.
- Course of Hematology and Hemostasis of graduate students, conducted by the Acute Hospital Interzonal "Dr. Gregorio Alfaro." Biochemical College of the Province of Buenos Aires, 08/19/1985.
Topic: thromboelastography.
- Refresher Course and systematization Hematological Pathology, conducted by the Society of Hematology Argentina, Buenos Aires 06/30/1986.
Topic: Testing for the study of platelet function, bleeding time, platelet adhesiveness in vitro, ristocetin cofactor, circulating platelet microaggregates, degranulation studies: labeling with mepacrine.
- Refresher Course conducted by the Department of Haemostasis and Thrombosis. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine.
Subject: Participation of platelets in the clotting mechanism. Platelet coagulant activity. Platelet Factor 3, Platelet Factor 4 and ß-Thromboglobulin: biochemistry and clinical interest. Detection of circulating platelet microaggregates: determination and clinical application. Laboratory control of oral anticoagulant treatment. Ristocetin Cofactor: determination and clinical usefulness. Von Willebrand disease: classification. Platelet specific proteins. From 1979 to 1993.
- Refresher Course conducted by the Department of Haemostasis and Thrombosis. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine. Module 1: "von Willebrand Disease." From 1994 to present.
Topic: ristocetin cofactor in plasma and platelets: identification and clinical utility.
- IIHema Joint Meeting of the National Academy of Medicine and the Association Argentina Hemotherapy and Immunohematology. Practical Module "Flow Cytometry." Bs As, 11.27.1998.
Topic: Determination of ristocetin cofactor in plasma, platelets, and cryoprecipitate.
- Haemostasis Module Career Specialist Clinical Biochemistry, Haematology area, Faculty of Pharmacy and Biochemistry, UBA, 06/08/2000.
Topic: Ristocetin Cofactor of: determining biochemical and clinical implications "
- Professional Development Course 2000. "Coagulation and Platelets: technical aspects." Argentina Technicians Association Clinical Laboratory. 14/7/2000.
Topic: Determination of ristocetin cofactor in plasma, platelets and cryoprecipitate: determination and clinical usefulness.
- Theoretical and Practical Course of Hemostasis and Thrombosis by the Department of Haemostasis and Thrombosis, 2002. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine. Module: Methodology applied to the study of thrombotic risk markers.
Topic: Determination of ristocetin cofactor in plasma, platelets and cryoprecipitate: determination and clinical usefulness.
- theoretical and practical course "Update in Hemostasis and Thrombosis" by the Department of Haemostasis and Thrombosis, 2003. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine.
Topic: Ristocetin cofactor activity: aggregation with ristocetin.
ristocetin cofactor activity in cryoprecipitate and concentrates of FVIII / VWF.
- theoretical and practical course "Update in Hemostasis and Thrombosis" by the Department of Hemostasis and Thrombosis, 16-23-5-2005. Module 1: Alterations in coagulation, with a total of 1:30 pm Chair. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine.
Topic: von Willebrand Factor. Von Willebrand disease and its variants.
ristocetin cofactor activity in plasma and platelets. Aggregation with ristocetin.
ristocetin cofactor activity in cryoprecipitate and concentrates of FVIII / VWF.
search techniques for mutation screening for VWD 2 N.
- theoretical and practical course organized by the Career Specialist in Clinical Biochemistry, Haematology area, 05.31.2006, Haemostasis Module "von Willebrand disease, diagnosis and classification."
- Speaker at the Second Scientific Meeting of the Argentina Society of Hematology on "Importance Molecular diagnosis of mutations in Hematology ", 27/04/1907.
Theme: "von Willebrand disease: molecular biology, detection of mutations."
- theoretical and practical course "Update in Hemostasis and Thrombosis" by the Department of Haemostasis and Thrombosis, 13 to 14/9/2007. Module III: Von Willebrand factor, with a total of 10 hours lecture. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine.
Topic: von Willebrand Factor. Von Willebrand disease, variants.
ristocetin cofactor activity in plasma and platelets. Aggregation with ristocetin. Biology
molecular vo Willebrand disease.
search techniques for mutation screening for VWD 2 N.
- Race Course organized by the Specialist in Clinical Biochemistry, Haematology area, 04.15.2008, Haemostasis Module "von Willebrand Factor, generalities. Von Willebrand disease, diagnosis and classification. Test ristocenina Cofactor of plasma and platelets. Aggregation with ristocetin. VWD molecular biology techniques, screening, sequencing.
PARTICIPATION IN RESEARCH PROJECTS
charge of sub-projects in the projects led by Dr Mary A. Lazzari described below: -1995
PID 332360092. PICT
0173 -1997 (PIP 4640/96). "Pared vascular, leukocytes, platelets and von Willebrand factor, its intervention
thrombosis." Project No. 05-00000-00706
ANPCyT -1998. "Physiology of platelets, leukocytes and endothelial cells mediated
cell interactions and von Willebrand factor." -1999
PIP CONICET Project No. 02066. "The von Willebrand factor. Its pathophysiological interactions.
von Willebrand disease"
-2000 ANPCyT Project PICT No. 8875. "Pathophysiology of humoral and cellular interactions in the vascular wall
." -2002
ANPCYT Project No. 0511270. Topic: "Structural alterations Factor von Willebrand endothelial cells produced by normal maternal plasma and purified proteases. Most common mutations and polymorphisms in the ADAMTS 13. " -2004
ANPCyT Project No. 25434. Topic:. ADAMTS-13: isolation, quantification in platelets (RT-PCR), pathologies and assessment models "in vivo" - "in vitro". -2004
PIP CONICET Project No. 6026. Topic: VWF and ADAMTS-13, molecular biology, quantification of mRNA in platelets. Similarity and difference in placental endothelium. Evaluation in human pathologies and animal models.
AWARDS OBTAINED
- Award of Masnatta Arini Elsa ", given by National Academy of Medicine. Buenos Aires, October 1988.
Topic: platelet disorders and the complex FVIII / von Willebrand factor in myelo-proliferative syndromes. Meschengieser S, Blanco A, Maugeri N, Woods A, Bermejo E, Dupont J, Fernandez J, Pieroni G, Lazzari MA
- Reach the Developing World Travel Grant. Awarded by the International Society on Thrombosis and Haemostasis. XIX Congress SSC and 49 ° Annual Meeting.
work won: Woods AI, Meschengieser SS, Blanco AN, Lazzari MA. Incidence of major haemorrhage related to surgery in type 1 von Willebrand's disease. XIX Congress and 49th Annual SSC Meeting. International Society on Thrombosis and Haemostasis. Birmingham, UK. jul12-18, 2003.
- Second Award Research Grant to ORT 2003 "Chemical ORT Project Human Genome" (95 points) with the theme "Diagnosis of von Willebrand disease (VWD) type 2N by genotypic and phenotypic techniques."
- Research Grant to ORT 2004 "Project Human Genome ORT Chemistry" with the theme "Diagnosis of von Willebrand disease (VWD) type 2N by genotypic and phenotypic techniques."
- Research Grant to ORT 2005 "Molecular Biology and Proteomics 2005" with the theme: VWF and ADAMTS-13: molecular biology.
- Grant for Research ORT 2006 in Molecular Biochemistry and Proteomics with the theme: Diagnosis of von Willebrand disease (VWD) type 2N by phenotypic and genotypic techniques.
- Research Grant to ORT 2007 in Molecular Biochemistry and Proteomics with the theme: "Study of ADAMTS-13 genotype in patients with thrombotic microangiopathy"
- Research Grant to ORT 2008 in Molecular Biochemistry and Proteomics with the theme: Diagnosis of von Willebrand disease (VWD) type 2N by phenotypic and genotypic techniques.
-
COLLABORATION IN BOOK CHAPTERS
Techniques Hemostasis and Thrombosis. Cooperative Group Latin American Hemostasis and Thrombosis, 2nd Edition. Editors: C. Campos Guerra, L. Kordich, JC Sanchez Avalos, O. Vidal.
- Detection of circulating platelet microaggregates: identification, interpretation and clinical utility.
- Ristocetin Cofactor: identification, interpretation and clinical utility.
Technical Manual. Institute of Hematologic Research "Mariano R. Castex", National Academy of Medicine. Editors: Luis J Bergner, Maria A Lazzari.
- Detection of circulating platelet microaggregates: detection, interpretation and clinical utility.
- Ristocetin Cofactor: identification, interpretation and utility clinic.
- platelet coagulant activity.
Foundations for the practical use in the laboratory of haemostasis. Argentinean Cooperative Group
Haemostasis and Thrombosis, 1 edition, 2003. Editor in Chief: Lucía Kordich. Chapter
Factor VIII: von Willebrand Factor:
- Ristocetin cofactor activity (VWF: RCo) in plasma. P. 295.
- intraplatelet von Willebrand factor. Ristocetin cofactor activity (VWF: RCo). P. 305. Chapter
Platelets Study Methodology:
- Determination of circulating platelet microaggregates. P. 124. VARIOUS
- Member of the Latin American Cooperative Group Haemostasis and Thrombosis (CLAHT) since 1980.
- Member of the Argentinean Cooperative Group of Haemostasis and Thrombosis (CAHT) since 1981.
- Concurrency to Henry Beaufour Research Institute, Paris, France, from 15 to 25/07/1987, which discussed various research projects with its Director, Dr. Eduardo Pirotski, concerning the effect of PAF and BN 52021 (its specific inhibitor) on the release of antiactivadores antiplasmin and plasminogen activator (PAI-1) of platelets.
- Secretary of the International Symposium on Thrombosis. Buenos Aires, Buenos Aires, 11/13 April 1988.
DIAGNOSIS OF VON WILLEBRAND DISEASE (VWD) type 2N FOR TECHNICAL Phenotype and genotype of the authors The project will Institute of Hematologic Research "Mariano R. Castex" of the National Academy of Medicine
INVESTIGATING:
Mary A. Lazzari. Assistant Director of the Institute of Hematologic Research "Mariano R. Castex. National Academy of Medicine. CONICET. Adriana I.
Woods. CONICET. Leticia
Keller. Biotechnology, National Academy of Medicine. INTRODUCTION
von Willebrand disease (vWD) is the most common human bleeding disorder (1) (up to 1% in some populations) and is produced by changes in factor von Willebrand (VWF), whose mode of inheritance is autosomal, although acquired cases may arise related to lymphoma and myeloproliferative syndromes. VWD can be divided into three main categories: type 1 and 3 are quantitative deficiencies and type 2 are qualitative deficiencies of vWF molecule. Type 3 is the least common, has very low levels or absent vWF and is inherited in a recessive, in type 2, although the protein may be present, is functionally defective and sub-classified into 4 groups: type 2A, 2M, 2B and 2N (2) with genotypic and phenotypic characteristics different for each subtype.
type 2N vWD (also called variant Normandy) is characterized by have low levels of Factor VIII (FVIII) compared with normal or subnormal levels of vWF and is inherited in a recessive (3). So far, we found that these patients were homozygous or double heterozygous for a series of mutations located between the domains D 'and D3 (4).
stabilization domain D 'requires disulfide bonds and cysteine \u200b\u200b(5), but some authors suggest that other amino acid residues (aa), are also required to maintain the functional conformation of this site. It suggests that the potential induced conformational change Asp116Asn mutation located in domain D3 may explain the loss of binding capacity of vWF to FVIII. It could also affect the formation of disulfide bonds intercadena involved in binding of vWF dimers to form multimers. A subset of type 2N vWD type 2N coheredó mutation in one allele and a mutation of type 1 or type 3 on the other allele. The expression of mutated recombinant vWF confirmed that a single aa substitution produces a decrease in the FVIII binding capacity (6). OBJECTIVES
• Detection of patients with von Willebrand disease type 2N phenotypic and genotypic techniques. Exons 17 to 24 DNA of patients with vWD for the diagnosis of vWD 2N to find known mutations and polymorphisms. Search
new mutations and polymorphisms in exons 25 to 28, and in the domain D3, they would be able to produce conformational changes in the domain D 'which alter the binding of vWF to FVIII.
estimate the prevalence in our variant within the vWD Normandy. Calculate the allele frequencies of different polymorphisms in different exons of the vWF to compare with those recorded in the database of vWF of different ethnic groups.
Using the technique of Conformation-Sensitive Gel Electrophoresis (CSGE) as a method of screening by heteroduplex formation, in order to detect if there was any alteration in a particular exon. In the case of detecting an altered electrophoretic pattern of exons in the CSGE, sequencing will make them for identification. EXPECTED RESULTS
The results of DNA sequencing of patients will allow us to relate their clinical symptoms with molecular alterations, to determine the prevalence of this variant in our population and find new polymorphisms and mutations.
METHODOLOGY AND WORK PLAN
Selection of samples for phenotypic screening methods:
1. Determination vWF antigen (vWf: Ag) by immunoelectrophoresis according to Laurell technique with anti-vWF polyclonal antibody (DAKO A082) in 1% agarose.
2. Determination of FVIII one stage method, using as a reagent, FVIII-deficient plasma.
3. Ristocetin Cofactor determination. Ristocetin agglutination of platelets in the presence of vWF, through a platelet membrane receptor, glycoprotein Ib, the high molecular weight multimers are required for this interaction while small multimers would be inactive in this regard. This determination is performed by the technique of Macfarlane.
4. Binding method FVIII to vWF. Are chosen patients with decreased levels of FVIII (VN: 0.5-1.5 U / dl) and normal or subnormal vWF: Ag (VN: 0.5-1.5 U / dl) and vWF: RCo (VN: 0.5-1.5 U / dl). The binding capacity of FVIII to vWF was determined using an ELISA previously described (7). The rationale is to incubate the reference samples and patients in polystyrene tubes coated with polyclonal anti-vWF which will join the complex FVIII / vWF samples. Eliminates the endogenous FVIII with CaCl2 and incubated the tubes with purified FVIII commercial. The FVIII bound is determined by the chromogenic substrate technique.
Grouping of patients with link mild or greatly diminished.
Based on the results of the prior art, establishing a ratio between the binding of vWF to FVIII of the patient and their level of vWF: Ag Up to a value of 0.8 is considered a normal link, below it, is considered mild or greatly diminished. DNA extraction
white blood cells.
Blood samples must be collected with EDTA to 2%. It used a commercial kit (Wizard Genomic DNA Purification Kit from Promega). The technique was performed following the manufacturer's instructions.
reaction polymerase chain (PCR).
Once extracted DNA must be the same for PCR amplification. This reaction carried out using 0.2 uL of Taq polymerase 0.5 U / mL, 1 g of DNA, 100 pM of each primer appropriate to the mutation being studied, 200 mmol / L of each deoxynucleotide triphosphate (dNTP) and magnesium chloride.
The following primers have been tested and allow the proper amplification of relevant exons: Exon 17
Primers: forward: 5 'GTG AGT GAG GCG GCA ATA G 3'
reverse: 5 'GAG CGT GGG TCT GAA CGA G 3 '
Exon 18 primers: forward: 5' GAC AAG AGT CCA GAA GGT GTG AGG 3 '.
reverse: 5 'CCT AAG GCC TAC AAA GGG GAA ACT 3'.
Exon 19 primers: forward: 5 'GGG AGA TTT TCA GTG ACT GTC 3'.
reverse: 5' GTG CAC CCT CAC TCC ACC CGC 3'.
Exón 20 Primers: forward: 5' GAT GGG CCC CTC AAA CCC AAG GTG 3'.
reverse: 5' CTC ACC TGC ACC AGA ACG TAC TGG 3'.
Exón 21 Primers: forward: 5' TTC TGG TCT GGT GAG AGC CAG TGG 3'.
reverse: 5' GCT TGG AAA TGT TCC TAT TAA GTG 3'.
Exón 23 Primers: forward: 5' TTC CCC TGA GCC GGA GAG GAT GCT 3'.
reverse: 5' GGA CAT TCC AGG AAG CAA GCT CTA 3'.
Exón 24 Primers: forward: 5'ATA ACC CCA GTT TGA CCA GCT GAC 3'.
reverse: 5'TCA CAC TCT GTG TCC ATA CCT CCA 3'.
Condiciones de la PCR para los exones 18, 19, 20, 21, 22, 23 y 24:
94ºC; 0:30min [94°C;0:30 min/58 ° C, 1:00 min/72 ° C, 1:00 min] x 40 cycles, 72 ° C, 10:00 min.
For exon 17 the temperarure of annealing (TA) is 56 ° C.
For exons 25, 26, 27 and 28 will design specific primers for amplification and test its operation.
DNA sequencing of patient samples.
To isolate the amplified fragment for the exon of interest, purify the PCR product by specific purification columns. After purification, PCR will be a second call sequencing reaction with specific primer (forward or reverse) plus DNA sequencing kit in order to amplify a single strand of DNA. The conditions of the sequencing reaction are: 94 ° C, 3:00 min [94 ° C, 30 ° C seg/50; 15seg/60 ° C, 4:00 min] x 25 cycles. Purified products of the sequencing reaction by precipitation with: absolute ethanol, 3 M sodium acetate, pH 4.8 and DNase free sterile water. The purified product was resuspended in TSR (template supression reagent) and proceed to the sequencing of the samples in the ABI Prism 310 Perkin Elmer.
CSGE (Conformation Sensitive Gel Electrophoresis).
The CSGE is used to detect mutations or polymorphisms based on the property that the change single nucleotide may alter the electrophoretic mobility of the PCR product under mild denaturing conditions (8). Under these conditions the DNA molecules have a stable three dimensional conformation depends on the sequence of nucleotides. Therefore, the single nucleotide change produces a conformational change that can be detected as an increase in the differential migration of homoduplex and heteroduplex in a polyacrylamide gel electrophoresis.
Statistics.
will use the Fisher test or chi square depending on n we obtain and consider a value of <>
significant if
