Person:
Bautista Santa Cruz, José Manuel

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First Name
José Manuel
Last Name
Bautista Santa Cruz
URI
https://hdl.handle.net/20.500.14352/80584
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Bioquímica y Biología Molecular
Area
Bioquímica y Biología Molecular
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End date: 2009 × Item Type: Publication ×

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    Synchronous culture of Plasmodium falciparum at high parasitemia levels
    (Nature Protocols, 2009) Radfar, Azar; Méndez, Dario; Moneriz, Carlos; Linares Gómez, María; Patricia Marín-García; Puyet Catalina, Antonio; Díez Martín, Amalia; Bautista Santa Cruz, José Manuel
    This protocol describes a method for preparing cultures of Plasmodium falciparum synchronized at any intraerythrocytic stage. Using this method, around 60% parasitized cells may be obtained. On the basis of Trager and Jensen's original continuous culture method, our approach relies on the use of fresh human blood not older than 2 weeks, a low hematocrit between 0.8 and 1.5%, a starting frozen inoculum of 10% ring-stage parasitemia, human serum replaced with AlbuMAX I and alternating sorbitol and Percoll synchronization methods to shorten the cycle window to 4–6 h and reduce sorbitol toxicity. From our synchronized high parasite density cultures, 3–5 ml of infected red blood cells can be obtained in 1 week, corresponding to 1.2 mg of total parasite protein per ml of harvested culture. On the basis of the variables parasitemia and packed cell volume, we provide an equation to accurately calculate the amount of complete medium required every 24 h corrected for the cycle stage and capacity of the culture flask. Ten days suffice to complete the protocol from a frozen stock of parasites.
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    Altered Nucleotide Receptor Expression in a Murine Model of Cerebral Malaria
    (The Journal of Infectious Diseases, 2009) Marín- García, Patricia; Sánchez‐Nogueiro, Jesús; Díez Martín, Amalia; León- Otegui, Míriam ; Linares Gómez, María; García Palencia, María Del Pilar; Bautista Santa Cruz, José Manuel; Miras Portugal, María Teresa
    In cerebral malaria, the most severe complication of malaria, both neurotransmission mechanisms and energy metabolism are affected. To understand how metabolic changes modify neurotransmission, we examined P2 receptor expression in a murine model of cerebral malaria. Quantitative polymerase chain reaction experiments revealed that parasite deposition was greatest in the cerebellum, compared with other areas of the brain, suggesting a correlation between brain parasitemia and loss of control of movement. Infected mice showed modified patterns of expression of P2 receptor subtype messenger RNA (mRNA), depending on both the specific purinergic receptor and the cerebral region analyzed. Immunohistochemical studies indicated altered levels of protein expression by these receptors in infected brains and, in some cases, a pattern of expression different from that noted in control mice. These differences in both the amount of mRNA and the protein distribution of P2 receptors observed in the different brain sites in infected mice suggest an important role for P2 receptors in either provoking cerebral damage or conferring neuroprotection.