Clinical Trial: Hemolysis in Patients With Hereditary Spherocytosis (HS)

Study Status: Completed
Recruit Status: Unknown status
Study Type: Interventional

Official Title: Oxidative Stress Contributes to Hemolysis in Patients With Hereditary Spherocytosis (HS) and Can be Ameliorated by Fermented Papaya Preparation (FPP)

Brief Summary: In the present study the investigators are going to explore the oxidative status of HS-RBC and its contribution to hemolysis

Detailed Summary:

The oxidative status of cells, which is determined by the balance between pro-oxidants, such as the reactive oxygen species (ROS), and antioxidants, is a major regulator of cellular functions. Impaired balance between pro- and antioxidants causes oxidative stress which may result in oxidation of proteins, lipids and DNA with the final outcome of premature cell aging and apoptosis [1,2]. Oxidatively stressed red blood cell (RBC) have been observed in various congenital and acquired hemolytic anemias, including thalassemia, sickle cell anemia, congenital dyserythropoietic anemia, G6PD deficiency and paroxysmal nocturnal hemoglobinuria (PNH) as well as in myelodysplastic syndrome (MDS). Although the primary etiology is different in these anemias, oxidative stress mediates several of their pathologies, mainly hemolysis [3].

Hereditary Spherocytosis (HS) is a genetic disorder of the RBC skeleton with primary deficiency in spectrin, ankyrin-1, band 3 or protein 4.2 associated with chronic hemolytic anemia [4]. Secondary protein deficiencies resulting from oxidative stress are often observed and may be involved in the clinical manifestations of the disease [5].


Sponsor: Wolfson Medical Center

Current Primary Outcome: ROS, reduced glutathione (GSH) and lipid peroxides will be measured in RBC [ Time Frame: year ]

ROS, reduced glutathione (GSH) and lipid peroxides will be measured in RBC following incubation with with 100 μM 2'-7'-dichlorofluorescin diacetate, 40 μM [1-(4-chloromercuryphenyl-azo-2-naphthol)] and fluor-DHPE, respectively for ROS [8] and with mercury orange for GSH [9]. After being washed twice, the cells will be resuspended in PBS and analyzed by flow cytometry .


Original Primary Outcome: Same as current

Current Secondary Outcome: Hemolysis [ Time Frame: year ]

Hemolysis will be assayed by suspending 3 ml of packed RBC in PBS or in the autologous plasma and overnight incubation in the presence of various concentrations of antioxidants such as fermented papaya preparation (FPP)


Original Secondary Outcome: Same as current

Information By: Wolfson Medical Center

Dates:
Date Received: September 12, 2010
Date Started: September 2010
Date Completion: September 2011
Last Updated: September 13, 2010
Last Verified: September 2010