scholarly journals Limited Plasmodium sporozoite gliding motility in the absence of TRAP family adhesins

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Konrad Beyer ◽  
Simon Kracht ◽  
Jessica Kehrer ◽  
Mirko Singer ◽  
Dennis Klug ◽  
...  

Abstract Background Plasmodium sporozoites are the highly motile forms of malaria-causing parasites that are transmitted by the mosquito to the vertebrate host. Sporozoites need to enter and cross several cellular and tissue barriers for which they employ a set of surface proteins. Three of these proteins are members of the thrombospondin related anonymous protein (TRAP) family. Here, potential additive, synergistic or antagonistic roles of these adhesion proteins were investigated. Methods Four transgenic Plasmodium berghei parasite lines that lacked two or all three of the TRAP family adhesins TRAP, TLP and TREP were generated using positive–negative selection. The parasite lines were investigated for their capacity to attach to and move on glass, their ability to egress from oocysts and their capacity to enter mosquito salivary glands. One strain was in addition interrogated for its capacity to infect mice. Results The major phenotype of the TRAP single gene deletion dominates additional gene deletion phenotypes. All parasite lines including the one lacking all three proteins were able to conduct some form of active, if unproductive movement. Conclusions The individual TRAP-family adhesins appear to play functionally distinct roles during motility and infection. Other proteins must contribute to substrate adhesion and gliding motility. Graphical Abstract

2021 ◽  
Author(s):  
QinQin Yu ◽  
Matti Gralka ◽  
Marie-Cécilia Duvernoy ◽  
Megan Sousa ◽  
Arbel Harpak ◽  
...  

AbstractDemographic noise, the change in the composition of a population due to random birth and death events, is an important driving force in evolution because it reduces the efficacy of natural selection. Demographic noise is typically thought to be set by the population size and the environment, but recent experiments with microbial range expansions have revealed substantial strain-level differences in demographic noise under the same growth conditions. Many genetic and phenotypic differences exist between strains; to what extent do single mutations change the strength of demographic noise? To investigate this question, we developed a high-throughput method for measuring demographic noise in colonies without the need for genetic manipulation. By applying this method to 191 randomly-selected single gene deletion strains from the E. coli Keio collection, we find that a typical single gene deletion mutation decreases demographic noise by 8% (maximal decrease: 81%). We find that the strength of demographic noise is an emergent trait at the population level that can be predicted by colony-level traits but not cell-level traits. The observed differences in demographic noise from single gene deletions can increase the establishment probability of beneficial mutations by almost an order of magnitude (compared to in the wild type). Our results show that single mutations can substantially alter adaptation through their effects on demographic noise and suggest that demographic noise can be an evolvable trait of a population.


2015 ◽  
Vol 22 (5) ◽  
pp. 895-906 ◽  
Author(s):  
Mark A. McCormick ◽  
Joe R. Delaney ◽  
Mitsuhiro Tsuchiya ◽  
Scott Tsuchiyama ◽  
Anna Shemorry ◽  
...  

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4136-4136
Author(s):  
Barbara J. Bryant ◽  
Julie A. Hopkins ◽  
Susan F. Leitman

Abstract Donors of apheresis blood components are routinely evaluated with a complete blood count (CBC) at the time of each donation. In otherwise healthy donors, recurrent low mean corpuscular volume (MCV) values (< 80 fL) in the presence of an acceptable hemoglobin (≥ 12.5 gm/dL) could be due to iron deficiency or to an hemoglobinopathy, such as alpha thalassemia trait or a beta chain variant trait. Iron deficiency in repeat blood donors may warrant treatment with oral iron supplementation, whereas donors with hemoglobinopathies in the absence of iron deficiency do not need treatment. Pre-donation samples for CBC (Cell-Dyn 4000, Abbott) were obtained from all apheresis donors donating platelets, plasma, granulocytes, lymphocytes, and monocytes. MCV values <80 fL were electronically flagged via a donor database module for review by medical staff. Donors with MCV ≤ 80 fL on two or more occasions were evaluated for iron deficiency and the presence of hemoglobinopathies. CBC, ferritin, serum iron, transferrin, percent transferrin saturation, and hemoglobin electrophoresis were performed at the time of a subsequent donation. Iron deficiency was defined as values below the reference range for ferritin or transferrin saturation. Alpha thalassemia trait was presumed if the red blood cell count was elevated, no variant hemoglobins were detected by electrophoresis, and the ferritin, percent transferrin saturation, serum iron, and transferrin levels were all within normal ranges. In a one-year period, 25 of 1333 healthy apheresis donors had a low MCV on more than one occasion. Donors with low MCV were more likely to be African American (AA) (12 of 25, 48%) or Asian (2 of 25, 8%) compared with donors without a low MCV (AA 193 of 1308, 15%; Asian 37 of 1308, 3%). Iron deficiency was present in 60% (15 of 25) of the low-MCV donors: 36% (9) had isolated iron deficiency, 20% (5) had iron deficiency with probable alpha thalassemia trait, and 4% (1) had hemoglobin C trait with coexistent iron deficiency. Hemoglobinopathy without concomitant iron deficiency was found in 40% (10 of 25) of the low-MCV donors and included 24% (6) with presumed alpha thalassemia trait, 4% (1) with hemoglobin S trait and single gene deletion alpha thalassemia trait (hemoglobin S concentration 34%), 4% (1) with hemoglobin S trait and double gene deletion alpha thalassemia trait (hemoglobin S concentration 28%), 4% (1) with hemoglobin Lepore trait, and 4% (1) with hemoglobin G-Philadelphia trait with at least a single gene deletion alpha thalassemia trait (hemoglobin G-Philadelphia concentration 36%). Although the combination of MCV, hemoglobin, and red cell count available from the routine CBC were often helpful in discriminating iron deficiency from hemoglobinopathy, the frequent coexistence of both processes resulted in a need for further laboratory evaluation, both before and after iron repletion, to confirm the diagnosis. In a sample of American repeat apheresis donors, iron deficiency is present in the majority with recurrent low MCV values and hemoglobin levels ≥ 12.5 gm/dL. Concurrent hemoglobinopathy is also commonly present but may not be easily recognized in the setting of iron deficiency. The MCV is a useful screening tool to detect iron deficiency in a repeat blood donor population, however low MCV values should be further investigated in the blood donor setting to determine if iron replacement therapy is indicated.


Yeast ◽  
2011 ◽  
Vol 28 (5) ◽  
pp. 349-361 ◽  
Author(s):  
Katsunori Yoshikawa ◽  
Tadamasa Tanaka ◽  
Yoshihiro Ida ◽  
Chikara Furusawa ◽  
Takashi Hirasawa ◽  
...  

2009 ◽  
Vol 5 (1) ◽  
pp. 335 ◽  
Author(s):  
Natsuko Yamamoto ◽  
Kenji Nakahigashi ◽  
Tomoko Nakamichi ◽  
Mihoko Yoshino ◽  
Yuki Takai ◽  
...  

2017 ◽  
Vol 16 (1) ◽  
Author(s):  
Aisyah Mohamed Rehan ◽  
Mohammad Izwan Enche Othman ◽  
Nor Munirah Mohd Amin ◽  
Intan Azura Shahdan ◽  
Hanani Ahmad Yusof@Hanafi

Introduction: Streptococcus pneumoniae (S. pneumoniae) is a gram-positive diplococci belonging to the genus Streptococcus and it is a well-studied pathogenic bacterium. Pneumococcal diseases such as otitis media, pneumonia, sepsis and meningitis caused by pathogenic strains of S. pneumoniae still brought significant mortality and morbidity worldwide. The pathogenicity of S. pneumoniae is exerted by various virulence factors and one of it is the enzyme hyaluronate lyase. Hyaluronate lyase plays a major role in the invasive capability of S. pneumoniae. Its mechanism of action and crystallographic structure have been determinedbut its regulatory mechanism is still poorly understood. Drawing connections between the nutritional behaviour and invasive property of S. pneumoniae, CodY regulator is hypothesized as a potential hyaluronate lyase regulator. This work was aimed to construct CodY deficient mutant of S. pneumoniae to form foundational work for the study of CodY regulatory effect on hyaluronate lyase. Materials and method: A single gene-deletion method was chosen to create CodY mutant construct containing aphA-3 gene (encoding kanamycin resistance protein) replacement cassette by employing recombinant fusion PCR method. A single band DNA product at the expected size of 2,563 bp after recombinant fusion PCR was visualized on agarose gel electrophoresis. This indicated that the linear recombinant amplicon has been successfully constructed. The recombinant amplicon was transformed into S. pneumoniae cells of Malaysian local clinical isolate by employing the competence stimulating peptide (CSP-1) to enhance and induce natural competence. Results: No viable CodY mutant was evident post-transformation. The negative transformation result was postulated to be due to the essential nature of CodY regulator. Conclusion: This work can provide basis for recombinant fusion PCR method in designing and constructing single gene deletion construct. However, further work is needed to identify the regulatory agent of hyaluronate lyase.


2021 ◽  
Author(s):  
T. L. Westerman ◽  
M. K. Sheats ◽  
J. R. Elfenbein

During enteric salmonellosis, neutrophil generated reactive oxygen species alter the gut microenvironment favoring survival of Salmonella Typhimurium. While the type-3 secretion system-1 (T3SS-1) and flagellar motility are potent Salmonella Typhimurium agonists of the neutrophil respiratory burst in vitro, neither of these pathways alone are responsible for stimulation of a maximal respiratory burst. In order to identify Salmonella Typhimurium genes that impact the magnitude of the neutrophil respiratory burst, we performed a two-step screen of defined mutant libraries in co-culture with human neutrophils. We first screened Salmonella Typhimurium mutants lacking defined genomic regions and then tested single gene deletion mutants representing particular regions under selection. A subset of single gene deletion mutants were selected for further investigation. Mutants in four genes, STM1696 (sapF), STM2201 (yeiE), STM2112 (wcaD), and STM2441 (cysA), induced an attenuated respiratory burst. We linked the altered respiratory burst to reduced T3SS-1 expression and/or altered flagellar motility for two mutants (ΔSTM1696 and ΔSTM2201). The ΔSTM2441 mutant, defective for sulfate transport, formed aggregates in minimal media and adhered to surfaces in rich media, suggesting a role for sulfur homeostasis in regulation of aggregation/adherence. We linked the aggregation/adherence phenotype of the ΔSTM2441 mutant to biofilm-associated protein A and flagellins and hypothesize that aggregation caused the observed reduction in the magnitude of the neutrophil respiratory burst. Our data demonstrate that Salmonella Typhimurium has numerous mechanisms to limit the magnitude of the neutrophil respiratory burst. These data further inform our understanding of how Salmonella may alter human neutrophil antimicrobial defenses.


Author(s):  
Inés Portillo-Calderón ◽  
Miriam Ortiz-Padilla ◽  
Jose Manuel Rodríguez-Martínez ◽  
Belen de Gregorio-Iaria ◽  
Jesús Blázquez ◽  
...  

Abstract Objectives To explore the effect of combining defects in DNA repair systems with the presence of fosfomycin-resistant mechanisms to explain the mechanisms underlying fosfomycin heteroresistance phenotypes in Enterobacteriaceae. Materials and methods We used 11 clinical Escherichia coli isolates together with isogenic single-gene deletion mutants in the E. coli DNA repair system or associated with fosfomycin resistance, combined with double-gene deletion mutants. Fosfomycin MICs were determined by gradient strip assay (GSA) and broth microdilution (BMD). Mutant frequencies for rifampicin (100 mg/L) and fosfomycin (50 and 200 mg/L) were determined. Using two starting inocula, in vitro fosfomycin activity was assessed over 24 h in growth (0.5–512 mg/L) and time–kill assays (64 and 307 mg/L). Results Strong and weak mutator clinical isolates and single-gene deletion mutants, except for ΔuhpT and ΔdnaQ, were susceptible by GSA. By BMD, the percentage of resistant clinical isolates reached 36%. Single-gene deletion mutants showed BMD MICs similar to those for subpopulations by GSA. Strong mutators showed a higher probability of selecting fosfomycin mutants at higher concentrations. By combining the two mechanisms of mutation, MICs and ranges of resistant subpopulations increased, enabling strains to survive at higher fosfomycin concentrations in growth monitoring assays. In time–kill assays, high inocula increased survival by 37.5% at 64 mg/L fosfomycin, compared with low starting inocula. Conclusions The origin and variability of the fosfomycin heteroresistance phenotype can be partially explained by high mutation frequencies together with mechanisms of fosfomycin resistance. Subpopulations should be considered until clinical meaning is established.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Takaaki Horinouchi ◽  
Tomoya Maeda ◽  
Hazuki Kotani ◽  
Chikara Furusawa

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