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Color Plates

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Image of COLOR PLATE 1 (Chapter 1)

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COLOR PLATE 1 (Chapter 1)

Appearance of by light microscopy after staining.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 2 (Chapter 2)

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COLOR PLATE 2 (Chapter 2)

Gene Pages on PlasmoDB.The summary view (upper left) provides a tabulation of the various forms of information available and scrolls to reveal highlights of general interest, such as curated annotations (and comments from the research community, which may also be entered by the user), DNA and protein features, transcript and protein expression data, information on protein structure, reagents, and publications. Insets illustrate for the DHFR-TS gene (PFD0830w) (i) a list of orthologs in other species, (ii) mapped protein motifs and features, (iii) transcript profiling data, including information on both abundance and inductionrepression, based on both photolithographic and glass slide microarrays, and (iv) the gene model(s), sequence, and information in local chromosomal DNA contexts.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 3 (Chapter 3)

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COLOR PLATE 3 (Chapter 3)

Illustrations summarizing the structural organizations of the major erythrocytic stages of , including a group of free and two invading merozoites (A), ring (B), trophozoite (C), schizont at the eight dividing nucleus (D), and late (E) developmental stages, and a mature macrogametocyte (F), shown with part of the surrounding RBC.Accompanying each schema is an example of a Giemsa-stained parasite from a film preparation of approximately the same stage. In panel F, three gametocytes are shown; from left to right, a mature (stage V) macrogametocyte and microgametocyte and an immature (stage IV) macrogametocyte with typical sharp ends (the oat-grain form).AP, apical prominence; Circl, circular cleft;Dmves, double membrane vesicle; Fvac, food vacuole; Hz, hemozoin; IMC, inner membrane complex; Mitoch, mitochondrion; Mz, merozoite; PM, plasma membrane; Pvac, parasitophorous vacuole;RER,rough endoplasmic reticulum;STIC, sexual-stage intraerythrocytic tubular compartment; ves, vesicle.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 3 (Chapter 3)

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COLOR PLATE 3 (Chapter 3)

Illustrations summarizing the structural organizations of the major erythrocytic stages of , including a group of free and two invading merozoites (A), ring (B), trophozoite (C), schizont at the eight dividing nucleus (D), and late (E) developmental stages, and a mature macrogametocyte (F), shown with part of the surrounding RBC.Accompanying each schema is an example of a Giemsa-stained parasite from a film preparation of approximately the same stage. In panel F, three gametocytes are shown; from left to right, a mature (stage V) macrogametocyte and microgametocyte and an immature (stage IV) macrogametocyte with typical sharp ends (the oat-grain form).AP, apical prominence; Circl, circular cleft;Dmves, double membrane vesicle; Fvac, food vacuole; Hz, hemozoin; IMC, inner membrane complex; Mitoch, mitochondrion; Mz, merozoite; PM, plasma membrane; Pvac, parasitophorous vacuole;RER,rough endoplasmic reticulum;STIC, sexual-stage intraerythrocytic tubular compartment; ves, vesicle.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 3 (Chapter 3)

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COLOR PLATE 3 (Chapter 3)

Illustrations summarizing the structural organizations of the major erythrocytic stages of , including a group of free and two invading merozoites (A), ring (B), trophozoite (C), schizont at the eight dividing nucleus (D), and late (E) developmental stages, and a mature macrogametocyte (F), shown with part of the surrounding RBC.Accompanying each schema is an example of a Giemsa-stained parasite from a film preparation of approximately the same stage. In panel F, three gametocytes are shown; from left to right, a mature (stage V) macrogametocyte and microgametocyte and an immature (stage IV) macrogametocyte with typical sharp ends (the oat-grain form).AP, apical prominence; Circl, circular cleft;Dmves, double membrane vesicle; Fvac, food vacuole; Hz, hemozoin; IMC, inner membrane complex; Mitoch, mitochondrion; Mz, merozoite; PM, plasma membrane; Pvac, parasitophorous vacuole;RER,rough endoplasmic reticulum;STIC, sexual-stage intraerythrocytic tubular compartment; ves, vesicle.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 4 (Chapter 5)

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COLOR PLATE 4 (Chapter 5)

Comparison of the cRNA hybridizing intensity level by the Affymetrix array for the trophozoite (red) and schizont (green) stages. Each colored square represents the hybridization signal from a single 25-bp probe to the genome sequence.The intensity of the color indicates hybridization level for each stage.Yellow equal hybridization to both trophozoite and schizont stages, red indicates hybridization for the trophozoite stages but not for the schizont stage, and green indicates hybridization for the schizont stage but not for the trophozoite stage.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 5 (Chapter 5)

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COLOR PLATE 5 (Chapter 5)

Comparison of the two-microarray analysis using the robust k-mean algorithm (A) with the short oligonucleotide array (reproduced from [Le Roch et al., 2003] with permission from the publisher) or FTT analysis (B) with the long 70-mer array (reproduced from [Bozdech et al., 2003a] with permission from the publisher).

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 6 (Chapter 8)

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COLOR PLATE 6 (Chapter 8)

Light microscopy of Giemsa-stained mature schizonts (strain IT04) and invading merozoites (strain NF54). (a) Both intact (upper) and rupturing (lower) schizonts are shown. (b) Initial apically attached merozoite; (c) partially invaded merozoite; (d) almost or fully invaded merozoite.The . (strain NF54) images were provided by Gabriele Margos (KCL).The images were generated with an Axiocam camera and a Zeiss Axiophot microscope.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 7 (Chapter 8)

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COLOR PLATE 7 (Chapter 8)

Diagrammatic representation of an invading merozoite and moving junction. (a) A theoretical molecular view of the electron-dense tight moving junction, showing the intimately associated red blood cell membrane and merozoite pellicle, along with current predicted molecular players, as detailed in the text. Here, a novel single-headed merozoite myosin, which is linked to the IMC, pulls the merozoite into the red blood cell on a short filament of F-actin, connected via aldolase to a membrane-bound merozoite ligand, which in turn binds to a red cell receptor. (b) A partially invaded merozoite highlighting major structures, parasitophorous vacuole formation, and likely biochemical events as entry proceeds. Organellar secretion, membrane generation, and host cell membrane restructuring contribute to the formation of the PVM. Points at which enzymolysis may occur are indicated.The moving junction “embracing the merozoite” as it enters the erythrocyte is detailed as a stippled band.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 8 (Chapter 11)

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COLOR PLATE 8 (Chapter 11)

fructose-1,6-bisphosphate aldolase (1A5C) (A), L-LDH complexed with NADH and oxamate (1LDG) (B), phosphoglycerate kinase adenosine monophosphate (1LTK) (C), and triose phosphate isomerase coupled to substrate analog 3-phosphoglycerate (1M70) (D).

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 9 (Chapter 13)

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COLOR PLATE 9 (Chapter 13)

Model of erythrocyte DRM rafts and their enrichment in the malarial vacuolar membrane.The uninfected erythrocyte membrane contains a dynamic milieu of generalized lipid domains (gray spheres) and raft microdomains (pink spheres) containing various proteins.Some proteins partition heavily into raft domains (i.e., flotillins), while others are only minimally present in rafts (i.e., band 3 and Glut1). During malaria infection, merozoite-stage parasites invade erythrocytes to reside in a membrane-bound parasitophorous vacuole.The PVM becomes selectively cholesterol enriched, and 10 of the known raft proteins are internalized to the PVM (flotillin 1 and flotillin 2, Gs,2-AR,AQP1,Duffy, CD55, CD58, CD59, and scramblase). Most of the abundant erythrocyte membrane proteins are not internalized to the PVM (i.e., glycophorins A and C and cytoskeleton-associated band 3).The lower-left inset shows the perspective of the model,which depicts a whole infected erythrocyte and a magnified view through the plasma membrane and PVM of a malaria-infected erythrocyte. Because the PVM is formed by invagination of the plasma membrane,proteins that are cytoplasmically oriented in uninfected cells remain so upon infection; protein structures exposed to the extracellular space face the vacuolar space upon infection. 4.1, band 4.1; Gs, G-protein αs; 2-AR, β2-adrenergic receptor;AQP1, aquaporin 1.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 10 (Chapter 13)

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COLOR PLATE 10 (Chapter 13)

Maurer's clefts are intermediates in PfHRPII transport and are targeted by its histidine-rich sequences. Single optical section of trophozoite-infected cells expressing PfHRPIIGFP (A) and SSGFPHis154 to -327 (B). Cells were fixed, permeabilized, subjected to indirect immunofluorescence assays, and probed with antibodies to GFP (green) and a Maurer's cleft resident protein, PfSBP (red). Magnifications of the indicated boxes in A and B are shown as A′ and B′, respectively.The extent of colocalization is shown in yellow. In A′ and B′, arrows indicate GFPlabeled tubules or vesicles connecting clefts (arrowheads) to each other or the vacuolar parasite (P). Nuclei were stained with Hoechst (blue). Bar, 5 µm.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 11 (Chapter 14)

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COLOR PLATE 11 (Chapter 14)

Plastid division in .The dividing apicoplast is labeled with green fluorescent protein fused to the apicoplast protein ACP.The nuclei are stained with 4′,6′-diamidino-2- phenylindole (DAPI; blue), and the food vacuole is stained with the marker of acidic compartments, Lysotracker (red) (Molecular Probes). Although apicoplast division in is synchronous with nuclear division, the dividing apicoplast (green) in does not appear to be strictly associated with the six or more newly forming nuclei (blue) in the early schizont. By the end of schizogony, a new plastid must segregate with each new nucleus.The microscopy was performed by students of the Biology of Parasitism Course, Marine Biological Laboratory, Woods Hole, Mass., in 2002.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 12 (Chapter 18)

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COLOR PLATE 12 (Chapter 18)

Intersubunit bridges (A) of secondary structure of 16S and 23S rRNAs showing the features involved in intersubunit contacts (red). (B and C) Interface view of the 50S and 30S subunits with the bridges numbered. RNA-RNA contacts are shown in magenta; protein-RNA and protein-protein contacts are shown in yellow.A, P, and E indicate the three tRNAs (23S rRNA) or tRNA anticodon stem loops (16S rRNA) (Yusupov et al., 2001).

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 13 (Chapter 18)

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COLOR PLATE 13 (Chapter 18)

Structure of the center of the large ribosomal subunit of . (a) Space-filling representation of the 50S particle (RNA in white and protein in yellow) in complex with three tRNAs (Lazaro et al., 1991).The subunit has been split to reveal the tunnel and the peptidyl transferase site (boxed). The orientation is the crown view, with the L1 protein to the left and the L7-L12 stalk to the right. (b) Close-up view of the active site shows the P-site, A-site, and the transfer RNA carrying the elongating peptide.The N3 of A2486 (A2451) (light blue) is in proximity to the 3′ OH of the deacylated product, and the base of U2620 (U2585) (red) has moved near to the newly formed peptidyl ester link and the 3′ OH of dimethyl A76. (Reproduced with permission from Schmeing et al. [2002].)

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 14 (Chapter 18)

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COLOR PLATE 14 (Chapter 18)

Superposition of antibiotic-binding locations and hydrophobic crevices. Surface representation of the ribosome (grey contour) shows that many antibiotics (stick figures) interact in part with the A-site crevice (green contour, upper middle) or with the exit tunnel hydrophobic crevice (green contour, lower right).These antibiotics overlap the binding site of a P-site substrate (orange) or that of an A-site substrate (red). (Reprinted from the with permission of the publisher [Hansen et al., 2003].)

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 15 (Chapter 21)

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COLOR PLATE 15 (Chapter 21)

Temporal regulation of gene expression. (A) IE undergo several morphological and phenotypic alterations during the asexual life cycle, changes that correlate roughly with stages of gene and protein expression. Examination of Giemsa-stained IE from synchronized cultures indicates that ring-stage parasites occur immediately after merozoite invasion of the erythrocyte up to roughly 15 to 20 hpi.The pigmented trophozoite stage follows from between approximately 20 to 40 hpi, with segmenting and schizont stages developing between 40 to 48 hpi. (B) transcription is tightly synchronized within ring-stage parasites and is followed by translation of a single major gene encoding the PfEMP1 variant exposed on the IE surface. PfEMP1 is first detected on the IE surface from 16 to 18 hpi and correlates with the first detection of a cytoadherence phenotype. At this stage, the level of full-length RNA transcript decreases, and it is no longer detected in mature pigmented trophozoites. PfEMP1 protein synthesis presumably ceases at this point. Segmenting and/or schizont stages correspond with poorly detected phenotype in in vitro studies, possibly due to fragile nature of the cells. Both RNA and phenotype are difficult to assess during this stage. (C) Phenotype-independent gene expression of . Transcripts for this gene are present in all parasites regardless of their cytoadherent phenotype, at a very low level in ring stages and at a very high level in pigmented trophozoites (Kyes et al., 2003). It is unclear whether this protein is translated or what role it might have in cytoadherence; in 3D7, is a pseudogene. It was originally thought that this gene encoded a PfEMP1 mediating the CSA-binding phenotype, but current data show that another gene, , encodes a more plausible candidate (Salanti et al., 2003).

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 16 (Chapter 22)

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COLOR PLATE 16 (Chapter 22)

Immunofluorescence assay with a mouse anti-human IgM monoclonal antibody and Alexa488 secondary antibody to show nonimmune IgM (green) on the surface of a rosetting infected erythrocyte. Parasite nuclei are stained blue with 4′,6′-diamidino-2-phenylindole, and red blood cells are stained red with concanavalin A-tetramethyl rhodamine isocyanate. (Reproduced from the [Rowe et al., 2002b] with permission from the publisher.)

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 17 (Chapter 23)

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COLOR PLATE 17 (Chapter 23)

(A) Ribbon diagram of the overall structure of dihydrofolate reductase-thymidylate synthase (PfDHFR- TS).The two DHFR monomers are shown in red and pink, while the two TS monomers are shown in grey and blue. (B) The structure of the complex of pyrimethamine in the active site of a mutant enzyme harboring the S108N mutation. The asparagine side chain clashes with the Cl group of pyrimethamine, resulting in decreased binding affinity. (C) Comparison of enzymeinhibitor interactions at the active sites of wild-type (lighter model) and the quadruple mutant PfDHFR- TS, in stereo view. Both enzymes are shown as a complex with WR99210 (WR, in cyan) and NADPH (NDP, in magenta).The flexible tail of WR99210 allows binding in a conformation that is unaffected by the pyrimethamine-resistant mutations (N51I+C59R+S108N+I164L) labeled in red.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 18 (Chapter 23)

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COLOR PLATE 18 (Chapter 23)

Distribution of MFQ IC50 values by copy number (triangles) and codon mutations (diamonds). Isolates (diamonds) are arranged in ascending order of MFQ resistance. (Reprinted from [Price et al., 2004] with permission from the publisher.)

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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Image of COLOR PLATE 19 (Chapter 26)

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COLOR PLATE 19 (Chapter 26)

In situ hybridization of BAC clone 24D05 to the polytene chromosome complement of .BAC DNA was labeled with Cy3 from Amersham; polytene chromosomes were counterstained with Yoyo-1 from Molecular Probes.

Citation: Sherman I. 2005. Color Plates, In Molecular Approaches to Malaria. ASM Press, Washington, DC.
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