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Application of optical tweezer technology reveals that PfEBA and PfRH ligands, not PfMSP1, play a central role in Plasmodium-falciparum merozoite-erythrocyte attachment, Supporting Information
<p>This repository contains the dataset and analysis scripts associated with the upcoming publication titled <em>Application of optical tweezer technology reveals that PfEBA and PfRH ligands, not PfMSP1, play a central role in Plasmodium-falciparum merozoite-erythrocyte attachment</em>. The repository includes a comprehensive collection of data and scripts related to optical tweezer experiments, growth assays, qPCR data, and supplementary information. It is organized into several sections, each detailing different aspects of the study:</p> <ul> <li><strong>Growth Assays:</strong> Includes raw and processed data on parasitemia levels, invasion rates, and growth rate assays, along with corresponding Jupyter notebooks and Python scripts for data visualization (e.g., <code>GrowthAssayPlotlib.py</code>, <code>Plot GA1.ipynb</code>, and <code>GA2_df_melted.json</code>).</li> <li><strong>qPCR Data:</strong> Contains results from multiple qPCR runs, including quantification data for various samples, as well as analysis scripts and plotted results (<code>qpcr_plotbench.ipynb</code>, <code>qPCR_plotting.py</code>, etc.). Data files such as <code>.xlsx</code> and <code>.json</code> contain gene expression data and fold changes to NF54.</li> <li><strong>Optical Tweezer Experiments:</strong> Includes detailed results and plots from optical tweezer measurements of attachment forces, time dependence, and multiple merozoite attachments. Notebooks (<code>tweezer_plots.ipynb</code>, <code>Antibody_binding_assay_plots.ipynb</code>) and data files support these analyses.</li> <li><strong>Optical Tweezer Images</strong>: Includes images that were used to measure RBC diameters for deformation and force measurements in <code>.tiff</code> format.</li> <li><strong>Supplementary Information (SI):</strong> Provides additional data and visualizations, such as scatter plots of two stretched RBCs, time post-egress vs. detachment force, and antibody GIA flow data. The accompanying figures (e.g., <code>SupFig1d_egress time vs force_3D7.svg</code>, <code>SupFig5a_GIA.svg</code>) are provided as <code>.svg</code> files.</li> </ul> <p>This repository offers all necessary resources to replicate the findings, including the complete codebase, raw data, and graphical representations of results. Researchers are encouraged to explore the included notebooks and datasets for detailed insights.</p>
Fig. 4 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 4. Comparison of fully grown gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of Curruca communis (A, B), Haemoproteus sittae (unknown lineage) from the blood of Sittae europaea (C, D) and Haemoproteus dolniki (unknown lineage) from the blood of Fringilla coelebs (E, F). Note: the elongate and markedly attenuated nuclei of infected erythrocytes containing advanced gametocytes of the new species (A, B), which is not the case in other Haemoproteus parasites (C–F). All images are from type specimens of these species. Symbols are the same as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.
Fig. 7 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 7. High gametocytaemia of Haemoproteus palloris (lineage hWW1) from the blood of a willow warbler Phylloscopus trochilus. Note that young gametocytes are absent, and the parasitaemia consists exclusively of mature fully grown macro- and microgametocytes, indicating a synchronous parasite development and probable absence of recent maturation of tissue meronts, which are the only source of merozoites for young gametocyte development during Haemoproteus infections. Triangle arrowhead – macrogametocytes; triangle wide arrowheads – microgametocytes. Long arrows – parasite nuclei. Giemsa-stained preparations. Scale bar = 10 µm.
Fig. 2 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 2. Gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of its type host, the common whitethroat Curruca communis: A-H – macrogametocytes, I-L – microgametocytes. Note: the markedly attenuated width of infected erythrocytes (D–G) containing advanced gametocytes, compared to uninfected erythrocytes, and the presence of predominantly oval or elongate pigment granules in fully grown gametocytes (F–H, J-L). All images are from the hapantotype. Long arrows – parasite nuclei. Short arrows – vacuoles. Arrowheads – pigment granules. Short simple wide arrow – nucleolus. Long triangle wide arrows – unfilled spaces between erythrocyte nuclei and gametocytes. Giemsa-stained thin blood films. Scale bar = 10 µm.
Fig. 6 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 6. Bayesian Inference tree (A) based on partial (478 bp) cytb sequences of Haemoproteus angustus n. sp. (lineage hCWT7) and the 20 closest related Haemoproteus lineages. Bayesian posterior probabilities and Maximum Likelihood bootstrap values were indicated above and below nodes, respectively. For each lineage, representative GenBank accession numbers and MalAvi lineage codes (if available) are indicated as well as the most common bird host. The scale bar indicates the expected mean number of substitutions per site according to the model of sequence evolution applied. Images B and C show the Median-Joining DNA haplotype network of partial (478 bp) cytb sequences of H. angustus hCWT7 and the 20 closest related Haemoproteus lineages. The upper image (B) shows the host distribution, and the lower image (C) depicts the geographic distribution according to the United Nations geoscheme. Each circle represents a unique haplotype/lineage. The frequency of each lineage is indicated for all haplotypes with more than one record and roughly corresponds to the size of circles. Bars on branches indicate the number of substitutions between two haplotypes. Small white circles represent median vectors, which are hypothetical (often ancestral or unsampled) sequences required to connect existing haplotypes with maximum parsimony.
Fig. 3 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 3. Comparison of fully grown gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of Curruca communis (A–D), H. belopolskyi (lineage hHIICT1) from the blood of Hippolais icterina (E–H) as well as H. parabelopolskyi (hSYAT2) (I–L), H. pallidulus (hSYAT3) (M–P), H. homogeneae (hSYAT16) (Q–T) and H. majoris (hWW2) (U–X) from the blood of Sylvia atricapilla. Note: the markedly attenuated gametocytes of the new species (A–D), which are not present in other Haemoproteus parasites (E–X); the fully grown gametocytes of H. belopolskyi (F, H) and H. parabelopolskyi (J, L) are bigger than those of H. angustus n. sp.; the pigment granules are predominantly roundish and small in H. pallidulus (M–P); the fully grown gametocytes are small and do not reach poles of infected erythrocytes in H. homogeneae (R, T); the erythrocyte nuclei are displaced laterally by H. majoris (V, X) – all these feature are not characteristic of H. angustus sp. nov. Symbols are the same as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.
Fig. 1. Bayesian phylogenetic tree constructed using partial cytochrome b in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 1. Bayesian phylogenetic tree constructed using partial cytochrome b sequences of 61 lineages of Haemoproteus, 4 lineages of Plasmodium, and Leucocytozoon sp. lSISKIN2 as outgroup. Posterior probabilities higher than 0.8 are indicated close to the respective nodes. Red font indicates the parasite lineage described in this publication. Vertical bars (A–D) show groups of closely related lineages, which complete development and produce gametocytes only in non-passerines (A, D), both non-passerines and passerines (B), and only passerines (C). Blue font indicates Haemoproteus species, which develop in non-passerine avian hosts, which are indicated by symbols (● – Psittaciformes; ∎ - Coraciiformes; ▴ - Strigiformes; ◆ - Anseriformes; ★ - Charadriiformes; ♥ - Pelecaniformes; ⋄ - Piciformes; ⊠ - Sphenisciformes; Ω - Musophagiformes; § - Trochiliformes; Ψ – Falconiformes; Σ – Columbiformes; Φ - Galliformes). Lineage names were provided (according to MalAvi database), followed by parasite species names and sequence GenBank accession numbers.
Fig. 5. Haemoproteus angustus n in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis
Fig. 5. Haemoproteus angustus n. sp. (lineage hCWT7) gametogenesis (A–H) and ookinete development (I–L) in vitro: A, B – initial stages of rounding up of gametocytes after the exposure of infected blood to air; C, E – rounded up macrogametocyte (C) and microgametocyte (E); F – the exflagellation; G – microgamete; H – fertilization; I – initial stage of ookinete development; J – medium differentiated ookinete; K – nearly mature ookinete with a residual body; L – mature ookinete without residual body. Note: the presence of several small residual bodies in erythrocytes containing gametocytes, which were preparing to escape from infected erythrocytes (A-C, E) – a unique character of this species during the initial stage of gametogenesis. Short triangle wide arrows – residual bodies; triangle arrowhead – macrogamete; triangle wide arrowheads – microgametes; short barbed arrows – finger-like outgrowth; long barbed arrows – apical end of developing ookinete; simple wide arrowhead – residual body of ookinete. Other symbols as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.
Fig. 2. Macroscopic examination. A in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study
Fig. 2. Macroscopic examination. A pale enlarged liver (Li); B hyperplastic nodules on the surface of the large spleen (S), inflated kidneys (K), and hypostatic congestion of left lung (Lu).
Fig. 5 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study
Fig. 5. Maximum-likelihood tree analysis based on partial cytb gene sequences (433 bp) of Haemoproteus from the studied sequence (linage BUTBUT15) and other sequences obtained from MalAvi database. Numbers on the branches indicate the percent of replicates that reproduced the topology for each clade. The percentages of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. Black circle indicates sequence obtained from the study.
Fig. 1 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study
Fig. 1. Gametocytes of haemosporidians (arrows) from the blood of the common buzzards. A-C mature macrogametocytes of L. buteonis in fusiform host cells; A host cell nucleus is not distorted but displaced laterally; B&C host cell nucleus is lateral and flattened; D young gametocyte of Haemoproteus. Giemsa stained thin blood films. Scale bar = 10 μm.
Fig. 4 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study
Fig. 4. Megalomeront of Haemoproteus sp. (linage BUTBUT15) from the kidney of a common buzzard. A&B same megalomeront of different magnifications (arrows). The structure is covered with a semi-thick capsule-like wall and contained highly eosinophilic irregularly-shaped cytomeres which have merozoites. Note that the host cell nucleus is not visible inside or close to megalomeront. Scale bars = 200 μm (A); 80 μm (B).
Fig. 3 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study
Fig. 3. Meronts of Haemoproteus sp. (linage BUTBUT15) from the lung of a common buzzard. A-D different shapes of pulmonary meronts (arrows). Each meront is surrounded by a thin wall and contained numerous basophilic round to oval merozoites. Note that the host cell nucleus is not visible in meronts. Scale bar = 40 μm.
Fig. 3 in Exo-erythrocytic development of Leucocytozoon parasites (Haemosporida, Leucocytozoidae) in song thrushes Turdus philomelos
Fig. 3. Tissue and blood stages of Leucocytozoon parasites in naturally infected song thrushes Turdus philomelos (individual no. 4, cytochrome b lineage lSTUR1), found in hematoxylin-eosin (H&E) stained (a-g, i), chromogenic in situ hybridization (CISH) treated (h and inserts a-c, e, i) histological sections. Early (a), advanced (b, c) and mature (d) meronts in the kidneys. Megalomeront-like structure (e-h) located close to a heart blood vessel (white asterisk) within the fat tissue (black asterisk). Leucocytozoon gametocytes in the kidneys (i). Simple black arrowheads: meronts. Simple white arrowhead: cytomeres (visible as darker stained nuclei surrounded by lighter stained clefts). Black triangle arrowhead: megalomeront-like structure. Short black triangle arrow: capsule-like wall. Short white triangle arrow: host cell nucleus in the capsule-like wall. Short simple white arrows: nuclei of infected host cells. Simple thin black arrows: gametocytes. Scale bars 20 μm, unless indicated otherwise.
Fig. 2 in Exo-erythrocytic development of Leucocytozoon parasites (Haemosporida, Leucocytozoidae) in song thrushes Turdus philomelos
Fig. 2. Tissue stages of Leucocytozoon parasites in naturally infected song thrushes Turdus philomelos, found in hematoxylin-eosin (H&E) stained (a, b, e, f) and chromogenic in situ hybridization (CISH) treated (insert in b, and c, d) histological sections. Early meronts in the kidneys of individual no. 1 (cytochrome b lineage lTUPHI14) (a), and advanced meronts in the kidneys of individual no. 2 (lSTUR1) (b) and no. 5 (co-infection of lSTUR1 and lTUPHI13) (e, f). Meronts labelled by a Leucocytozoon-specific (Leuco18S) probe in the kidneys (insert b), and lungs of individual no. 2 (lSTUR1) (c) and individual no. 3 (lSTUR1) (d). Tissue stage developing in the Bowman capsule of a renal corpuscle (e). Leucocytozoon blood stage labelled by a Leucocytozoon-specific probe (c). Simple black arrowheads: meronts. Simple white arrowheads: cytomeres (visible as darker stained nuclei surrounded by lighter staining clefts). Short simple black arrow: nucleus of Leucocytozoon blood stage. Scale bars 20 μm.
Fig. 1 in Exo-erythrocytic development of Leucocytozoon parasites (Haemosporida, Leucocytozoidae) in song thrushes Turdus philomelos
Fig. 1. Mature gametocytes of Leucocytozoon dubreuili in roundish host cells from the blood of a song thrush Turdus philomelos (individual no. 1, cytochrome b lineage lTUPHI14). Macrogametocyte (a). Microgametocyte (b). Short simple white arrows: nuclei of infected host cells. Short simple black arrow: parasite nucleus. Long simple white arrows: vacuoles. Long simple black arrow: volutin granules (small purplish dots). Methanol-fixed and Giemsa-stained. Scale bar 10 μm.
Data from: Transmission pathways and spillover of an erythrocytic bacterial pathogen from domestic cats to wild felids
Many pathogens infect multiple hosts, and spillover from domestic to wild species poses a significant risk for spread of diseases that threaten wildlife and humans. Documentation of cross-species transmission, and unravelling the mechanisms that drive it, remains a challenge. Focusing on co-occurring domestic and wild felids, we evaluate possible transmission mechanisms and evidence of spillover of 'Candidatus Mycoplasma haemominutum' (CMhm), an erythrocytic bacterial parasite of cats. We examine transmission and possibility of spillover by analysing CMhm prevalence, modeling possible transmission pathways, deducing genotypes of CMhm pathogens infecting felid hosts based on sequences of the bacterial 16S rRNA gene, and conducting phylogenetic analyses with ancestral state reconstruction to identify likely cross-species transmission events. Model selection analyses suggest both indirect (i.e., spread via vectors) and direct (i.e., via inter-specific predation) pathways may play a role in CMhm transmission. Phylogenetic analyses indicate that transmission of CMhm appears to predominate within host species, with occasional spillover, at unknown frequency, between species. These analyses are consistent with transmission by predation of smaller cats by larger species, with subsequent within-species persistence after spillover. Our results implicate domestic cats as a source of global dispersal and spillover to wild felids via predation. We contribute to the emerging documentation of predation as a common means of pathogen spillover from domestic to wild cats, including pathogens of global conservation significance. These findings suggest risks for top predators as bioaccumulators of pathogens from subordinate species.
Erythrocyte sedimentation rate in the evolution of covid -19 patients attended at the CLAS health center, Nuevo Lurin-2021
<p>Abstract</p> <p><strong>Background:</strong> To determine the values of the erythrocyte sedimentation rate in the evolution of patients with COVID -19 with treatment attended at the CLAS Health Center, Nuevo Lurín.</p> <p><strong>Methods:</strong> The research was applied with an observational and descriptive method. The approach was quantitative, applied and cross-sectional. The study population was 13,000 patients in the CLAS center in Nuevo Lurin. After applying the inclusion and exclusion criteria, there were only 45 male and female patients attended at the CLAS Health Center in Nuevo Lurin between February and June 2021, to whom the capillary method was applied in order to determine the values of the erythrocyte sedimentation rate in the evolution of COVID-19 patients. The data collection instrument was a record card.</p> <p><strong>Results:</strong> It was compared that between male and female sex, there are statistically significant differences between the 45 COVID-19 patients evaluated. Higher values were identified in 7 male patients, while in women there was 1 patient with elevated values of erythrocyte sedimentation rate. This shows that there is a higher incidence of erythrocyte sedimentation rate in males.</p> <p><strong>Conclusion: </strong>Values of erythrocyte sedimentation rate were determined in 45 patients with COVID-19 evolution with treatment attended at the CLAS Health Center in Nuevo Lurin and confirming that erythrocyte sedimentation rate evolves significantly in COVID-19 patients.</p>
DNA replication dynamics during erythrocytic schizogony in the malaria parasites Plasmodium falciparum and Plasmodium knowlesi
<p>Malaria parasites are unusual, early-diverging protozoans with non-canonical cell cycles. They do not undergo binary fission, but divide primarily by schizogony. This is a mode of replication involving asynchronous production of multiple nuclei within the same cytoplasm, culminating in a single mass cytokinesis event. The rate and efficiency of parasite replication is fundamentally important to malarial disease, which tends to be severe in hosts with high parasite loads. Here, we have studied for the first time the dynamics of schizogony in two human malaria parasite species, <em>Plasmodium falciparum</em> and <em>Plasmodium knowlesi</em>. These differ in their cell-cycle length, the number of progeny produced and the genome composition, among other factors. Comparing them could therefore yield new information about the parameters and limitations of schizogony. We report that the dynamics of schizogony differ significantly between these two species, most strikingly in the gap phases between successive nuclear replications, which are longer in <em>P. falciparum</em> and shorter, but more heterogenous, in <em>P. knowlesi</em>. In both species, gaps become longer as schizogony progresses, whereas each period of active replication grows shorter. In both species there is also extreme variability between individual cells, with some schizonts producing many more nuclei than others, and some individual nuclei arresting their replication for many hours while adjacent nuclei continue to replicate. The efficiency of schizogony is probably influenced by a complex set of factors in both the parasite and its host cell.</p>
Within-lizard erythrocyte size variation
<p>Dataset associated to submitted article: To vary or not to vary: Is within-individual variation in erythrocyte size related to Plasmodium infection in a tropical lizard?<br> <br> Contains measurements of <em>A. gundlachi</em> examination season (season), snout to vent length (svl) , unique identifier (uid), lizard species (spp), sex (sex), <em>Plasmodium azurophilum</em> infection status (infectionstatus), mean erythrocyte area (erymeanarea), standard deviation of erythrocyte area (erysdarea). </p>
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