We are developing a new reagent for sickle cell screening, but we found following problem
Why the new cell transform to echinocytes?
We are facing new problem. The new reagent for sickle test has been adjusted and we found optima concentration. However when aplied to fresh red blood cell, imediately the RBC transform to echinocytes. Is it the ´´Holly leaf shape``.Why it happens?
The pH is around 7,2. No pH range afect the experiment.
Caracteristically it appears like a toothed cell that presents / displays on its surface small progressions rounded like bombs or vesicles of uniform size and symmetrically distributed that with the MEB, they resemble a sea urchin.
The transformation of the dystrocyte-equinocito can be produced "in vitro" by exposing the red cells to various inducing agents such as: suspension of erythrocytes in saline, interaction with glass surfaces or lytic agents (lysolecithin, saponins, among others); These changes are reversible if the inducing agent is removed before loss of the membrane material occurs. When they are induced experimentally they occur in fractions of seconds, without compromise of the water content nor loss of membrane fragments.
As has been said, ATP is required, among other things, for the control of active cation transport pumps; In particular, the lack of ATP alters the Ca ++ output, thus increasing its intracellular content, which in turn leads to a selective loss of K +, water outflow and cell dehydration called the Gardos Phenomenon.
The active pumping of Ca ++ is critical for the red cell because high concentrations of this cation are associated with adverse physiological effects: at the micromolar levels, formation of equinocytes occurs as a consequence of the Gardos Phenomenon. At millimolar levels, the alteration compromises contractile proteins (spectrin and ankirin) and membrane phospholipids, generating equinoctic cells with membrane rigidity described as sphero-equinocytes representing the terminal stage of the Ca ++ overloaded cell. In summary the lack of ATP and the intracellular accumulation of Ca ++ are equino-cytogenic.
Echinocytes are seen in patients with: Uremia, defects in glycolytic metabolism (pyruvate kinase deficiency) and in some patients with microangiopathic anemia. They are common in neonates and are observed transiently after massive transfusions of stored blood (storage injury) and characterize the metabolic depletion of the senile red cell.
Thanks so much professor. May follow my project please.