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Schizonts

Schizonts are a stage in the life cycle of certain parasitic protozoans, particularly those that cause malaria.
Schizonts are the product of asexual reproduction within host cells, where the parasite undergoes multiple rounds of nuclear division to produce numerous daughter merozoites.
This schizont stage is crucial for the propagation of the infection within the host.
Understanding the biology and behavior of schizonts is essential for developing effective treatments and preventive strategies against malaria and other schizont-producing parasitic diseases.
Researchres can leverage PubCompare.ai's AI-driven platform to quickly locate the best protocols from published literature, preprints, and patents to optimize their schizont research, saving time and enhancing the quality of their work.

Most cited protocols related to «Schizonts»

Artemisinin Resistance Evaluation Protocol

Cited 62 times

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Publication 2013

artenimol Atmosphere Biological Assay BLOOD Cell Nucleus Erythrocytes Gentamicin Heparin Sodium Parasitemia Parasites Percoll Pharmaceutical Preparations Schizonts Sorbitol Sulfoxide, Dimethyl Thermal Plasma Trophozoite Volumes, Packed Erythrocyte

In vitro Culture and Genetic Manipulation of P. falciparum

Cited 36 times

P. falciparum parasites were grown in vitro in RPMI 1640 medium containing 0.5% w/v AlbumaxII as described^{42 (link)}. Parasites were synchronised by purifying schizont stages using a 70% Percoll gradient, before allowing reinvasion to occur, and followed by sorbitol treatment. All parasites used in this study were derived from the 3D7 clone, including the control 1G5DC strain^{13 (link)}. Plasmids were introduced into purified schizont stage parasites by electroporation using an Amaxa 4D-Nucleofector™ (Lonza) as previously described^{43 (link)}. For transfections with reporter plasmids to be maintained episomally 10 μg of DNA was electroporated and cultures were selected continuously with 2.5 μg/ml of blasticidin-S-HCl. For generation of marker-free, DiCre transgenic parasites 20–30 μg of CRISPR/Cas9 plasmids and 60 μg of linearised rescue plasmids were electroporated. Selection was applied one day after transfection with 5 nM WR99210 (a kind gift of Jacobus Pharmaceuticals) for four days. Following establishment of transgenic parasites, the cultures were treated with 1 μM 5-fluorocytosine (5-FC) provided as clinical grade Ancotil^® (MEDA) for one week before parasite cloning by limiting dilution. To induce DiCre recombinase mediated excision of DNA, early ring stage parasites were treated with rapamycin (stock solution was 200 μM in DMSO), before being washed in warm RPMI 1640 medium and returned to culture.

Knuepfer E., Napiorkowska M., van Ooij C, & Holder A.A. (2017). Generating conditional gene knockouts in Plasmodium – a toolkit to produce stable DiCre recombinase-expressing parasite lines using CRISPR/Cas9. Scientific Reports, 7, 3881.

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Publication 2017

Ancobon Animals, Transgenic blasticidin S Clustered Regularly Interspaced Short Palindromic Repeats Electroporation Parasites Percoll Pharmaceutical Preparations Plasmids Recombinase Schizonts Sirolimus Sorbitol Sulfoxide, Dimethyl Technique, Dilution Transfection

In Vitro Cultivation and Genetic Manipulation of Malaria Parasite P. falciparum

Cited 33 times

Asexual blood-stage cultures of P. falciparum clone 3D7 were maintained in vitro and synchronized according to standard procedures (Blackman, 1994 (link); Yeoh et al., 2007 (link)) in RPMI 1640 medium containing the serum substitute Albumax (Invitrogen). For introduction of transfection constructs, mature schizonts were enriched from highly synchronous cultures using Percoll (GE Healthcare) as described previously (Harris et al., 2005 (link)), and transfected by electroporation with 10 μg of circular plasmid DNA using the Amaxa 4D electroporator (Lonza) and the P3 Primary cell 4D Nucleofector X Kit L (Lonza) and program FP158, exactly as recently described for P. knowlesi (Moon et al., 2013 (link)). Selection for parasites harbouring the plasmid was performed by culture in medium containing 2.5 nM WR99210. Selection for parasites in which integration of transfected DNA into the genome had taken place was promoted by cycles of culture in the absence and presence of WR99210, as described previously (Harris et al., 2005 (link)). When integration was detected by diagnostic PCR, integrant clones were obtained by limiting dilution and maintained in medium containing WR99210. Parasite growth rates were assessed by microscopic examination of Giemsa-stained thin films at 2-day intervals, and expressed as percentage parasitaemia (percentage of parasitized erythrocytes in the population)

Collins C.R., Das S., Wong E.H., Andenmatten N., Stallmach R., Hackett F., Herman J.P., Müller S., Meissner M, & Blackman M.J. (2013). Robust inducible Cre recombinase activity in the human malaria parasite Plasmodium falciparum enables efficient gene deletion within a single asexual erythrocytic growth cycle. Molecular Microbiology, 88(4), 687-701.

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Publication 2013

BLOOD Clone Cells Diagnosis DNA, Circular Electroporation Erythrocytes Genome L Cells Microscopy Parasitemia Parasites Percoll Plasmids Schizonts Serum Stain, Giemsa Technique, Dilution Transfection

Microscopic analysis of P. vivax parasites

Cited 27 times

Thick and thin smears were routinely made for the pre- and post-CF11 filtration from each of the P. vivax isolates collected, prior to ex vivo drug sensitivity testing at the Shoklo Malaria Research Unit (SMRU), Mae Sod, Thailand. The pre- and post-CF11 filtration, thick and thin smears from 30 randomly selected isolates collected during 2008 were examined as follows. Parasitaemias were determined from the number of IRBC per ten 100× oil immersion fields (200 RBC per field) on the thin film. The percentage of early (ring-like parasites with a single chromatin dot) and mature (amoeboid-like cytoplasm or presence of haemozoin) asexual stages was determined from examining 200 parasites in the thick smears under 100× oil immersion. Due to their very low numbers in the pre- and post-filtration smears schizonts (parasites with 3 or more chromatin dots and haemozoin) were combined with the mature stage count. Gametocyte counts were too low for statistical comparison.
Parametric analysis of non-paired data was calculated using one-way analysis of variance. Non-parametric analysis of the paired data was performed using Wilcoxon or Friedman's tests and post-hoc analysis using Dunn's test (GraphPad Prism 5.01).
The clinical IRBC samples examined in this study were collected under the following ethical guidelines in the approved protocol OXTREC 027-05 (University of Oxford, Centre for Clinical Vaccinology and Tropical Medicine, UK)

Sriprawat K., Kaewpongsri S., Suwanarusk R., Leimanis M.L., Lek-Uthai U., Phyo A.P., Snounou G., Russell B., Renia L, & Nosten F. (2009). Effective and cheap removal of leukocytes and platelets from Plasmodium vivax infected blood. Malaria Journal, 8, 115.

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Publication 2009

Amoeba Chromatin Cytoplasm Filtration hemozoin Hypersensitivity Immersion Malaria Oil Fields Parasites prisma Schizonts

Microarray Analysis of Plasmodium falciparum

Cited 23 times

The microarray used in this study is based on long oligonucleotides (70 nt) designed against the PlasmoDB 4.4 release of the 3D7 reference P. falciparum genome (Hu et al. 2007 (link)). We reannotated the microarray against the 3D7 PlasmoDB 7.0 genome release, which resulted in changes in more than 700 probes, including removal of cross-reactive probes. Additional probes were excluded for the analysis of parasites genetically different from 3D7, because some probes fall within highly polymorphic regions of the genome (Supplemental Figs. S4, S10, S12; Supplemental Table S5; Supplemental Methods).
RNA was purified using the TRIzol method, and cDNA was synthesized by reverse transcription, purified, labeled, and hybridized essentially as previously described (Bozdech et al. 2003 (link)). Microarray images were analyzed using GenePix Pro 6.0 (Axon Instruments), with careful visual inspection to refine automatic spot definitions and to eliminate poor quality spots. All test samples (labeled with Cy5) were hybridized against a reference pool (labeled with Cy3), which consisted of a mixture of cDNA from 3D7-A and 3D7-B rings, trophozoites, and schizonts in equal amounts for samples from 3D7-derived parasite lines, or from D10, 7G8, and HB3 for all other samples. 3D7 was not included in the parentals comparison because it was hybridized against a different reference pool. For CGH analysis, sonicated gDNA of test parasite lines was labeled with Cy5 and hybridized against 3D7-A gDNA labeled with Cy3.

Rovira-Graells N., Gupta A.P., Planet E., Crowley V.M., Mok S., Ribas de Pouplana L., Preiser P.R., Bozdech Z, & Cortés A. (2012). Transcriptional variation in the malaria parasite Plasmodium falciparum. Genome Research, 22(5), 925-938.

Publication 2012

Axon Cross Reactions DNA, Complementary Exanthema Figs Genome Microarray Analysis Neutrophil Oligonucleotides Parasites Parent Reverse Transcription Schizonts trizol Trophozoite

Most recents protocols related to «Schizonts»

Isolation and Treatment of Synchronised 3D7 Schizonts

Tightly synchronised 3D7 schizonts expressing endogenously GFP-tagged GAPM2 (glideosome-associated protein with multiple membrane spans 2^{56 (link),57 (link)}) were isolated from a 10–20 mL culture (5–10% parasitaemia) using 60% Percoll ^{55 (link)}, washed twice in pre-warmed RPMI, and treated with 1 µM of the PKG-inhibitor compound 2 (provided by Dr. Mike Blackman, The Francis Crick Institute, UK)^{58 (link),59 (link)}, and incubated for 6 h. Segmented schizonts were washed once and then resuspended in pre-warmed RPMI without Albumax or phenol red but with the addition of 1 µM E64 (Sigma) to allow the parasitophorous vacuole to rupture but prevent rupture of the red blood cell membrane. This helped us identify very late stage schizonts in the SEM. Cells were kept warm and vitrified within an hour.

Ferreira J.L., Pražák V., Vasishtan D., Siggel M., Hentzschel F., Binder A.M., Pietsch E., Kosinski J., Frischknecht F., Gilberger T.W, & Grünewald K. (2023). Variable microtubule architecture in the malaria parasite. Nature Communications, 14, 1216.

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Publication 2023

Cells Erythrocyte Membrane Membrane Proteins Parasitemia Percoll Schizonts Vacuole

Parasite Life Cycle Visualization

Five preparations of P. falciparum gametocytes were generated of which six grids were imaged, five preparations of P. falciparum schizonts were generated of which six grids were imaged, two preparations of P. falciparum sporozoites were generated of which four grids were imaged and two preparations of P. berghei ookinetes were generated of which three grids were imaged.

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Publication 2023

Schizonts Sporozoites

Synchronizing Plasmodium Intraerythrocytic Cycle

Schizonts were harvested using 60% prewarmed Percoll^{55 (link)}, washed once with prewarmed RPMI, resuspended in 5% uninfected red blood cells in prewarmed RPMI and cultured for 3 h, until parasites egressed and reinvaded. Unruptured schizonts were lysed with 5% sorbitol. The remaining culture contained young rings with a 3-h synchronicity window.

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Publication 2023

Erythrocytes Parasites Schizonts Sorbitol

Parasite Membrane Protein Extraction

Schizont-stage parasites were lysed with 0.1% saponin and washed 3 times with ice-cold PBS. Parasite pellets were resuspended in ice-cold lysis buffer (1× PBS, 1% Triton X-114 [Thermo Scientific 28332], 2 mM EDTA, 1× protease inhibitors [Pierce A32955]) and incubated on ice for 30 min. Cell debris was removed by a 10-minute centrifugation at 16,000 × g, 4°C. Supernatant was transferred to a fresh Eppendorf tube, incubated for 2 min at 37°C to allow phase separation, and centrifuged for 5 min at 16,000 × g at room temperature. The top (aqueous) layer was transferred to another tube. The interphase was removed to avoid cross-contamination between the layers. The bottom (detergent) layer was resuspended in 1× PBS-0.2 mM EDTA to equalize the volumes of the two fractions. Both fractions were subjected to methanol-chloroform precipitation, resuspended in PBS containing 2× NuPAGE lithium dodecyl sulfate sample buffer, boiled for 5 min at 95°C, and analyzed by Western blotting. Quantification of band intensities was done using Image Studio Lite. The percentage of each GFP fusion protein that was membrane bound was calculated and normalized to the percentage of endogenous Atg8 in the membrane.

Walczak M., Meister T.R., Nguyen H.M., Zhu Y., Besteiro S, & Yeh E. (2023). Structure-Function Relationship for a Divergent Atg8 Protein Required for a Nonautophagic Function in Apicomplexan Parasites. mBio, 14(1), e03642-21.

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Publication 2023

Buffers Cells Centrifugation Chloroform Cold Temperature Detergents dodecyl sulfate, lithium salt Edetic Acid Interphase Methanol Parasites Pellets, Drug Protease Inhibitors Proteins Saponin Schizonts Tissue, Membrane Triton X-114

Parasitemia Measurement by Flow Cytometry

Ring-stage parasites at 5% to 10% parasitemia were washed twice in the culture medium to remove ATc and resuspended in the culture medium, and the hematocrit was adjusted to 2%. Parasites were divided into 2 cultures grown in the culture medium supplemented with 0.5 μM ATc or grown in the medium without ATc. The growth assays were continued for 4 reinvasion cycles. At the schizont stage of each cycle, parasitemia was measured by flow cytometry starting at 24 h (0 reinvasions) after ATc removal. Culture aliquots were incubated with 17 μM dihydroethidium (Thermo Fisher) for 15 min at room temperature (RT) to stain nuclei, and 50,000 events were analyzed on a BD Accuri C6 flow cytometer. Cultures were diluted with fresh culture medium with red blood cells at 2% hematocrit so that the parasitemia under the +ATc condition was 1% and the −ATc condition was diluted by the same factor; ATc was added as required. Aliquots of culture for Western blotting were taken at 24 h and 72 h (0 and 1 reinvasion) after ATc removal.

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Publication 2023

Biological Assay Cell Nucleus Culture Media dihydroethidium Erythrocytes Flow Cytometry Parasitemia Parasites Schizonts Stains Volumes, Packed Erythrocyte

Top products related to «Schizonts»

Albumax 2 by Thermo Fisher Scientific

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Albumax II is a supplement used in cell culture media to support the growth and proliferation of cells. It is a highly purified bovine serum albumin (BSA) product.

Percoll by GE Healthcare

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Percoll is a colloidal silica-based medium used for cell separation and gradient centrifugation. It is designed to provide a density gradient for the isolation and purification of cells, organelles, and other biological particles.

Rpmi 1640 medium by Thermo Fisher Scientific

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RPMI 1640 medium is a commonly used cell culture medium developed at Roswell Park Memorial Institute. It is a balanced salt solution that provides essential nutrients, vitamins, and amino acids to support the growth and maintenance of a variety of cell types in vitro.

Hypoxanthine by Merck Group

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Percoll by Merck Group

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Percoll is a colloidal silica-based density gradient medium used for the separation and purification of cells, organelles, and other biological particles. It is designed to create density gradients for the isolation of specific cell types or subcellular fractions through centrifugation.

Rpmi 1640 by Thermo Fisher Scientific

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RPMI 1640 is a common cell culture medium used for the in vitro cultivation of a variety of cells, including human and animal cells. It provides a balanced salt solution and a source of essential nutrients and growth factors to support cell growth and proliferation.

Albumax 1 by Thermo Fisher Scientific

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Albumax I is a cell culture supplement provided by Thermo Fisher Scientific. It serves as a serum replacement in cell culture media formulations.

Giemsa by Merck Group

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Giemsa is a type of stain used in microscopy for the differential staining of cells, tissues, and other biological samples. It is a widely used stain in various fields, including hematology, parasitology, and cytology. Giemsa stain primarily helps to visualize and differentiate cellular components, such as nuclei, cytoplasm, and specific organelles, based on their varying affinity for the stain.

Albumax by Thermo Fisher Scientific

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Magnetic activated cell sorting (macs) by Miltenyi Biotec

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MACS is a magnetic cell separation technology developed by Miltenyi Biotec. It enables the efficient isolation and enrichment of target cells from complex biological samples. The core function of MACS is to provide a gentle, reliable, and scalable method for cell separation and purification.

What are the different types of Schizonts?

Schizonts can be classified into several types based on the specific parasitic protozoan species they are associated with. The most well-known are the Plasmodium schizonts, which are the schizont stage of the malaria-causing Plasmodium parasites. Other types of schizonts include those found in Toxoplasma, Babesia, and Cryptosporidium species, among others. Each type of schizont has unique characteristics and behaviors that are important to understand for effective research and treatment.

How can PubCompare.ai help with Schizont research?

PubCompare.ai's AI-driven platfrom can be invaluable for Schizont research in several ways. First, it allows you to quickly screen through a large volume of published literature, preprints, and patents to identify the most relevant and effective protocols related to Schizonts. The platform's intelligent comparison feature can help you pinpoint critical insights, such as differences in protocol effectiveness, that enable you to choose the best option for reproducibility and accuracy in your work. This saves you time and enhances the quality of your Schizont research.

What are some common challenges in working with Schizonts?

One of the main challenges in working with Schizonts is their complex life cycle and behavior within host cells. Schizonts undergo multiple rounds of nuclear division, making them difficult to isolate and study. Additionally, Schizonts can exhibit variability in their characteristics depending on the specific parasite species and host environment. Proper experimental design and optimization of protocols are crucial to overcome these challenges and obtain reliable and reproducible results.

How can PubCompare.ai help researchers find the best Schizont protocols?

PubCompare.ai's advanced AI technology can be extremely helpful in locating the most effective and reproducible protocols for Schizont research. The platfrom's intelligent comparison features allow you to quickly sift through a large volume of published literature, preprints, and patents to identify the protocols that are most relevant to your specific research goals. By highlighting key differences in protocol effectiveness, PubCompare.ai enables you to choose the best option to ensure the accuracy and reliability of your Schizont studies, saving you valuable time and effort.

What are some practical applications of Schizont research?

Schizont research is crucial for developing effective treatments and preventive strategies against malaria and other parasitic diseases caused by Schizont-producing protozoa. Understanding the biology and behavior of Schizonts, such as their role in the parasite's life cycle and their interactions with host cells, can lead to the identification of new drug targets and the optimization of existing therapies. Morover, Schizont research can also contribute to the development of improved diagnostic tools and vaccines to enhance the prevention and control of these parasitic infections.

More about "Schizonts"

Schizonts are a crucial stage in the life cycle of certain parasitic protozoans, particularly those that cause malaria.
These intracellular structures are the result of asexual reproduction, where the parasite undergoes multiple rounds of nuclear division to produce numerous daughter merozoites.
This schizont stage is essential for the propagation of the infection within the host.
Understanding the biology and behavior of schizonts is crucial for developing effective treatments and preventive strategies against malaria and other schizont-producing parasitic diseases.
Researchers can leverage advanced platforms like PubCompare.ai to quickly locate the best protocols from published literature, preprints, and patents, optimizing their schizont research and enhancing the quality of their work.
In the lab, researchers often utilize various media and supplements to culture and study schizonts, such as Albumax II, Percoll, RPMI 1640 medium, Hypoxanthine, and Albumax I.
These tools help maintain the viability and growth of the parasitic cultures.
Additionally, techniques like Giemsa staining can be employed to visualize and analyze the schizont stage under the microscope.
By delving into the complexities of schizonts, scientists can uncover valuable insights that contribute to the development of novel therapies and preventive measures against malaria and other schizont-producing parasitic infections.
PubCompare.ai's AI-driven platform can be a powerful ally in this endeavor, helping researchers navigate the vast body of published literature and find the most relevant and reproducible protocols to optimize their schizont research, saving time and enhancing the quality of their work.