Histopathological, Immunohistochemical and Molecular Detection of Toxoplasma gondii in Organs and Tissues of Experimentally Infected Mice

Purpose: Toxoplasmosis is a multisystemic disease of Toxoplasma gondii, an obligate intracellular protozoan that can affect all vertebral organisms. The aim of this study is to determine the distribution rate histopathologically, immunohistochemically and molecularly with mouse experiments. Methods: In our study, T. gondii TR01 tachyzoites were injected intraperitoneally into Specific Pathogen Free, 3-4 weeks old, 17-18 gr healthy white male Swiss-Albino mice. Two mice were euthanized daily and the distribution of the parasite was determined in daily concentrations in blood, peritoneal fluid, liver, kidney, heart, lung, intestine and central nervous system sections. Results: As a result of the histopathological analyzes, mild necrosis and degeneration of hepatocytes were observed in the liver on the 1st day. Tachyzoites are common in central regions. Degeneration was observed in the cortical and corticomedullary tubular epithelium in the kidney. Slightly degeneration of cardiomyocytes was observed in the heart. Regional hyperemic capillaries were found in the lung. Degeneration of intestinal epithelial cells was observed, and necrosis was not observed in villi and glandular epithelium. In the CNS, cerebral cortical neurons were observed to be affected by degeneration and necrosis. On the 2nd day, degeneration and necrosis of hepatocytes in the liver were observed moderately compared to the previous day. Degeneration of cortical and medullary tubules was more common in the kidney. Mild necrosis and degeneration were observed in some villi epithelium in the intestine. Lesions on day 3th were as on day 2th of the experiment. Degeneration and necrosis were detected in the renal tubules. The lung was more hyperemic. Degeneration and necrosis were more prominent in the cortical neurons of the brain. Lesions on day 4th were the same as on days 2th and 3th of the experiment. On the 5th day, the parasite was found to be less than the degeneration and necrosis of the tubules in the kidney tissue. In immunohistochemical staining, reactions were mostly found in liver, kidney, and intestine, while relatively low levels were found in lung, heart, and brain. RT-PCR targeting the T. gondii B1 gene region molecularly was used. All tests were positive except day zero in RT-PCR. The CT value and the lower values indicate the areas with the highest density. On the first day and on the last day, it is mostly found in the liver, lung and peritoneal fluid. At least, it spread to the brain and heart. Conclusion: Factors such as host cell type, infection rate, susceptibility, route of administration, and host and tissue selection of the parasite affect the virulence of the T. gondii strain. Knowing which strain caused the infection may be useful in predicting the outcome of organ involvement and virulence. The death of all mice within 6 days in our study indicates that the T. gondii TR 01 strain is a highly aggressive and lethal agent for mice. Histopathological, immunological and PCR spread to the CNS was found to be minimal. This is thought to be because the infective agent is less able to cross the CSF barrier. Tissue cysts were not found in all tissues. T. gondii infects the liver first in all three studies and remains in high concentration until death. Low spread to the heart is also similar to other studies. Desquamation and leukocytosis were not observed in the lung alveoli in our study. The liver, kidney and peritoneal fluid are most affected, and the brain is the least affected. This situation informs us that the parasite invasion in the liver is intense and is in parallel with other studies. Necrosis was detected in all tissues except the intestine. Toxoplasma gondii has the potential to develop changes by spreading in tissues. In today's world where drug and vaccine studies are increasing rapidly, it is important to know the distribution of the parasite in the tissues and the changes it makes. In this context, it is a parasite that still needs to be investigated together with the parasite strains, which is mandatory and difficult to diagnose and treat

Polygenic burden in focal and generalized epilepsies

Rare genetic variants can cause epilepsy, and genetic testing has been widely adopted for severe, paediatric-onset epilepsies. The phenotypic consequences of common genetic risk burden for epilepsies and their potential future clinical applications have not yet been determined. Using polygenic risk scores (PRS) from a European-ancestry genome-wide association study in generalized and focal epilepsy, we quantified common genetic burden in patients with generalized epilepsy (GE-PRS) or focal epilepsy (FE-PRS) from two independent non-Finnish European cohorts (Epi25 Consortium, n = 5705; Cleveland Clinic Epilepsy Center, n = 620; both compared to 20 435 controls). One Finnish-ancestry population isolate (Finnish-ancestry Epi25, n = 449; compared to 1559 controls), two European-ancestry biobanks (UK Biobank, n = 383 656; Vanderbilt biorepository, n = 49 494), and one Japaneseancestry biobank (BioBank Japan, n = 168 680) were used for additional replications. Across 8386 patients with epilepsy and 622 212 population controls, we found and replicated significantly higher GE-PRS in patients with generalized epilepsy of European-ancestry compared to patients with focal epilepsy (Epi25: P = 1.64 710-15; Cleveland: P = 2.85 710-4; Finnish-ancestry Epi25: P = 1.80 710-4) or population controls (Epi25: P = 2.35 710-70; Cleveland: P = 1.43 710-7; Finnish-ancestry Epi25: P = 3.11 710-4; UK Biobank and Vanderbilt biorepository meta-analysis: P = 7.99 710-4). FE-PRS were significantly higher in patients with focal epilepsy compared to controls in the non-Finnish, non-biobank cohorts (Epi25: P = 5.74 710-19; Cleveland: P = 1.69 710-6). European ancestry-derived PRS did not predict generalized epilepsy or focal epilepsy in Japanese-ancestry individuals. Finally, we observed a significant 4.6-fold and a 4.5-fold enrichment of patients with generalized epilepsy compared to controls in the top 0.5% highest GE-PRS of the two non-Finnish European cohorts (Epi25: P = 2.60 710-15; Cleveland: P = 1.39 710-2). We conclude that common variant risk associated with epilepsy is significantly enriched in multiple cohorts of patients with epilepsy compared to controls-in particular for generalized epilepsy. As sample sizes and PRS accuracy continue to increase with further common variant discovery, PRS could complement established clinical biomarkers and augment genetic testing for patient classification, comorbidity research, and potentially targeted treatment

Genome-wide identification and phenotypic characterization of seizure-associated copy number variations in 741,075 individuals

Copy number variants (CNV) are established risk factors for neurodevelopmental disorders with seizures or epilepsy. With the hypothesis that seizure disorders share genetic risk factors, we pooled CNV data from 10,590 individuals with seizure disorders, 16,109 individuals with clinically validated epilepsy, and 492,324 population controls and identified 25 genome-wide significant loci, 22 of which are novel for seizure disorders, such as deletions at 1p36.33, 1q44, 2p21-p16.3, 3q29, 8p23.3-p23.2, 9p24.3, 10q26.3, 15q11.2, 15q12-q13.1, 16p12.2, 17q21.31, duplications at 2q13, 9q34.3, 16p13.3, 17q12, 19p13.3, 20q13.33, and reciprocal CNVs at 16p11.2, and 22q11.21. Using genetic data from additional 248,751 individuals with 23 neuropsychiatric phenotypes, we explored the pleiotropy of these 25 loci. Finally, in a subset of individuals with epilepsy and detailed clinical data available, we performed phenome-wide association analyses between individual CNVs and clinical annotations categorized through the Human Phenotype Ontology (HPO). For six CNVs, we identified 19 significant associations with specific HPO terms and generated, for all CNVs, phenotype signatures across 17 clinical categories relevant for epileptologists. This is the most comprehensive investigation of CNVs in epilepsy and related seizure disorders, with potential implications for clinical practice

The Synthesis, Self-Assembly and Self-Organisation of Polysilane Block Copolymers

Block copolymers containing polysilane blocks are unique in that the polysilane components possess electro-active properties and are readily photodegradable. This review will discuss and assess the two major approaches to the synthesis of polysilane block copolymers via pre-formed polymer chain coupling and living polymerisation techniques. The self-organisation of polysilane block copolymers and the morphologies adopted in thin films are reviewed. Amphiphilic polysilane-containing block copolymers self-assemble in solvents selective for one block and a number of examples are highlighted. The versatility of these materials is highlighted by recent significant applications including the preparation of hollow crosslinked micellar aggregates in aqueous solutions and in patterned thin film generation subsequently employed as templates for the growth of cell cultures and CaCO (3.

Conceptual Modelling for Simulation-Based Serious Gaming

In recent years several simulation-based serious games have been developed for mastering new business concepts in operations management. This indicates the high potential of simulation use for pedagogical purposes. Unfortunately, this potential is hardly reflected in simulation methodology. We consider this issue by identifying alternative demands game use of simulation sets for model building and application. Moreover, we propose a framework for conceptual modelling for simulation-based serious gaming, which addresses relevant issues in a systematic, step-wise manner. Use of the framework is illustrated by two case examples, highlighting simulation use for training and education respectively

Parametric and distribution-free bootstrapping in robust simulation-optimization

Most methods in simulation-optimization assume known environments, whereas this research accounts for uncertain environments combining Taguchi’s world view with either regression or Kriging (also called Gaussian Process) metamodels (emulators, response surfaces, surrogates). These metamodels are combined with Non-Linear Mathematical Programming (NLMP) to find robust solutions. Varying the constraint values in this NLMP gives an estimated Pareto frontier. To account for the variability of this estimated Pareto frontier, this contribution considers different bootstrap methods to obtain confidence regions for a given solution. This methodology is illustrated through some case studies selected from the literature