Genotyping of European Toxoplasma gondii strains by a new high-resolution next-generation sequencing-based method.

PURPOSE: A new high-resolution next-generation sequencing (NGS)-based method was established to type closely related European type II Toxoplasma gondii strains. METHODS: T. gondii field isolates were collected from different parts of Europe and assessed by whole genome sequencing (WGS). In comparison to ME49 (a type II reference strain), highly polymorphic regions (HPRs) were identified, showing a considerable number of single nucleotide polymorphisms (SNPs). After confirmation by Sanger sequencing, 18 HPRs were used to design a primer panel for multiplex PCR to establish a multilocus Ion AmpliSeq typing method. Toxoplasma gondii isolates and T. gondii present in clinical samples were typed with the new method. The sensitivity of the method was tested with serially diluted reference DNA samples. RESULTS: Among type II specimens, the method could differentiate the same number of haplotypes as the reference standard, microsatellite (MS) typing. Passages of the same isolates and specimens originating from abortion outbreaks were identified as identical. In addition, seven different genotypes, two atypical and two recombinant specimens were clearly distinguished from each other by the method. Furthermore, almost all SNPs detected by the Ion AmpliSeq method corresponded to those expected based on WGS. By testing serially diluted DNA samples, the method exhibited a similar analytical sensitivity as MS typing. CONCLUSION: The new method can distinguish different T. gondii genotypes and detect intra-genotype variability among European type II T. gondii strains. Furthermore, with WGS data additional target regions can be added to the method to potentially increase typing resolution.

A comprehensive guide to telocytes and their great potential in cardiovascular system.

Telocytes, a recently discovered type of interstitial cells, have a very distinctive morphology - the small cell body with long extensions, named telopodes. In our review, apart from introducing general aspects of telocytes, we focus on properties, functions and future potential of those cells in cardiovascular system. However, physiological functions of telocytes in cardiovascular system are still regarded as quite enigmatic. Previous studies claim that they play a role in organogenesis and regeneration, bioelectrical signalling, mechanoelectrical coupling, anti-oxidative protection, angiogenesis and regulation of blood flow. As well, they are presumably connected with the presence of blood-myocardium barrier and proper organisation of extracellular matrix. Moreover, there exists a significant link between the quantity of telocytes in tissue and numerous cardiovascular diseases such as: myocardial infarction, cardiomyopathies, systemic sclerosis, heart failure, atrial fibrillation, isolated atrial amyloidosis, myxomatous valve degeneration and hyperplastic consequences of vascular injury. Thanks to their unique properties, telocytes might be a breakthrough in treatment of cardiovascular diseases, as they may be effective in reversing effects of myocardial infarction. Telocytes also may play a major role in tissue engineering - they might be the key factor in creating stable and efficient vascular network in larger synthetic tissues or organs (Tab. 1, Fig. 3, Ref. 53).

Configurations of the circle of Willis: a computed tomography angiography based study on a Polish population.

The aim of the study was to investigate the distribution of the circle of Willis variants in Polish population by means of computed tomography angiography (CTA). The results were then analysed and compared with another study that used similar methods but that was carried out on an ethnically distinct population. Patients presenting with intracranial pathology were excluded from the initial study population. In total, 250 CTA belonging to 129 female and 121 male patients were reviewed. A modified classification system of the circle was proposed, which took into consideration the anterior and the posterior aspects of the circle individually. The typical variant of Willis's circle occurred in 16.80% of cases. The anterior and the posterior portions of the circle were normal in 47.20% and 26.80% of the patients respectively. As for the anterior part, lack of the anterior communicating artery was the most frequent abnormality (22.80%). Bilateral absence of posterior communicating arteries was the most common anomaly in the posterior part of the circle (29.20%). This type of anomaly was also the most common, when taking into consideration the entire circle (12.00%). There were statistically significant differences between the age groups and genders when considering the occurrence of an incomplete circle. Overall, a substantial proportion of patients manifested clinically important variants that were incapable of providing collateral circulation. Comparison with other imaging-based and cadaveric studies revealed noticeable differences, that may have resulted from the variable technical features of other studies or other factors such as the ethnical origins of the studied populations.

Variations in hepatic vascularisation: lack of a proper hepatic artery. Two case reports.

The blood supply of the liver and other abdominal organs plays a significant role during abdominal surgery. Knowledge of the most common patterns of vascularisation should be broadened and new anomalies of the celiac trunk and its branches dutifully reported. This paper presents two case reports which describe the lack of a proper hepatic artery. Case 1 describes the cadaver of a 64-year-old female in whom the right hepatic artery was observed to arise from the common hepatic artery and run behind the portal vein. The common hepatic artery was observed to be divided into three terminal vessels: the left hepatic artery, the gastroduodenal artery, and the right gastric artery. Case 2 describes the cadaver of a 75-year-old male with a liver that was supplied from 3 different sources: the left hepatic artery from the left gastric artery (which arose directly from the aorta), the right hepatic artery from the superior mesenteric artery, and the middle hepatic artery from the common hepatic artery - (branch of the hepato-splenic trunk). Moreover, the left inferior phrenic artery arose from the left hepatic artery.

The median nerve in the carpal tunnel.

A study of the variations of the course and branching pattern of the median nerve within the carpal tunnel were carried out on 60 wrists from 30 fresh cadavers autopsied in the Department of Forensic Medicine of Jagiellonian University Medical College. The results were compared with the literature. The study confirmed that the extraligamentous type of motor branch variation is most common. The transligamentous course of the nerve is of special importance: it is usually accompanied by hypertrophic muscle, and the nerve hidden within this muscle can easily be cut during transection of the retinaculum. The results proved the necessity of approaching the median nerve from the ulnar side when opening the carpal tunnel.