Benzylpenicillin is incubated with whole human blood. The degree of reactivity of 14C-benzylpenicillin to the components of this blood is determined via a Beckman LS-100C Liquid Scintillation Counter.The oxygen carrying capacities of penicillin treated adult normal and sickled hemoglobin are measured spectrophotometrically and compared to untreated adult normal and sickled hemoglobin.Ball State UniversityMuncie, IN 47306

The technique of ¹H spin echo n.m.r. has been used for the non-invasive study of enzyme catalysed ¹H/²H equilibrium isotope exchange at the C-2 position of lactate in suspensions of human erythroeytes. The intracellular environment of the enzymes involved in this exchange has been investigated by comparing the exchange properties of the enzymes in the intact cell with the properties they display in vitro. A study of the exchange of the lactate C-2 substituent with solvent, which is catalysed by a coupled system of four glycolytic enzymes, has teen used to examine the kinetic properties of the individual enzymes in vitro. Measurements of the exchange in the intact cell have been used to investigate the in situ kinetic properties of one of these enzymes, glyceraldehydephosphate dehydrogenase. Contrary to the conclusions of previous studies with the isolated enzyme in vitro, these measurements have shown that the enzyme is not rate determining for glycolytic flux in the human erythrocyte and that it is unlikely that it is bound to the cell membrane in situ. A study of ¹H/²H exchange between the C-2 positions of methyl labelled lactate molecules, catalysed by lactate dehydrogenase, has been used to investigate the in situ kinetic properties of this enzyme. Comparison of these properties with those it displays in vitro indicate that the free intracellular NAD(H) concentration in the erythrocyte is only approximately 10% of the total extractable concentration. A considerable fraction of the coenzyme must be bound, therefore, in the intact cell. This type of experiment should be widely applicable to a variety of tissues and possibly to different dehydrogenases. Theoretical aspects of bulk isotope exchange kinetics in multi-enzyme systems are examined and the effects of chemical flux, and of isotope effects, on the measurement of isotopic flux are considered. The advantages of the n.m.r. method over conventional radioactive tracer techniques are described. It is concluded that ¹H n.m.r. studies of ¹H/²H isotope exchange may be used to obtain information about the kinetic properties of enzymes in intact cellular systems. The technique should be a useful complement, therefore, to the currently more widely used n.m.r. methods employing the ³¹P and ¹³C nuclei and to other methods used for the non-invasive study of metabolism.

Severe anemia is the most prevalent life-threatening complication of malaria infection. In addition to destruction of red blood cells (RBC), decreased RBC production or erythropoietic suppression has been shown to contribute to malarial anemia. The mechanism of this suppression is unknown, but it is considered to be multifactorial since erythropoietic suppression can be observed in the presence of both inflammatory mediators and parasite-derived factors. Experiments presented in this thesis aimed at determining the role of host cytokines released in response to blood-stage malaria infection and parasite-derived factors in erythropoietic suppression underlying the development of malarial anemia. Pro-inflammatory cytokines released during malaria infection have been proposed to play a central role in erythroid suppression. To dissect the discrete roles of these cytokines in the processes leading to anemia, mice were treated with CpG-oligodeoxynucleotides (CpG-ODN) which, like malaria infection in humans and experimental mouse models, induces an acute type 1 pro-inflammatory response. CpG-ODN treatment induced anemia, which was associated with suppressed erythropoiesis and reduced RBC survival. Importantly, CpG-ODN-induced IFN-gamma was found to be the major factor mediating erythropoietic suppression but not decreased RBC survival. We also studied the roles of Th1, Th2 and anti-inflammatory cytokines produced in response to Plasmodium chabaudi AS infection in the development of erythropoietic suppression during blood-stage malaria. Signal transducer and activator of transcription (STAT)6, required for signaling of the Th2 cytokines IL-4 and IL-13, was shown to play a critical role in malarial anemia by inhibiting the proliferation and differentiation of erythroid cells. We also observed that suppressed erythropoiesis is a general feature in mice infected with various rodent Plasmodium species that differ in their clinical manifestations and immune responses. Since parasite-derived factors have been shown to contribute to malarial pathogenesis including anemia, the contribution of P. falciparum - and P. yoelii-derived products to erythropoietic suppression was investigated. Both Plasmodium-derived and synthetic hemozoin (Hz) suppressed the proliferation but not the maturation of erythroid progenitor cells in vitro. However, P. yoelii-derived Hz but not synthetic Hz induced transient anemia in mice. These findings provide novel insights into the complex interactions between the parasite and host immune system and the regulation of erythropoiesis during severe malarial anemia.

Diss. (sammanfattning) Uppsala : Univ., 2003. / Härtill 9 uppsatser.

Diss. (sammanfattning) Linköping : Univ., 2001. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Bilagan utgöres av sammanfattnig på svenska med titeln: Fysiologiskt svar på arbete hos varmblodiga travhästar i relation till total blodvolym. Härtill 5 uppsatser.

Thesis (Ph. D. in Molecular Physiology and Biophysics)--Vanderbilt University, May 2006. / Title from title screen. Includes bibliographical references.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Feb. 18, 2009). Includes bibliographical references.

Thesis (Ph.D. (Tropical Medicine))--Mahidol University, 2000.