Novel Porcine Circoviruses in View of Lessons Learned from Porcine Circovirus Type 2-Epidemiology and Threat to Pigs and Other Species

Porcine circovirus type 2 (PCV2) plays a key role in PCV2-associated disease (PCVAD) etiology and has yielded significant losses in the pig husbandry in the last 20 years. However, the impact of two recently described species of porcine circoviruses, PCV3 and PCV4, on the pork industry remains unknown. The presence of PCV3 has been associated with several clinical presentations in pigs. Reproductive failure and multisystemic inflammation have been reported most consistently. The clinical symptoms, anatomopathological changes and interaction with other pathogens during PCV3 infection in pigs indicate that PCV3 might be pathogenic for these animals and can cause economic losses in the swine industry similar to PCV2, which makes PCV3 worth including in the differential list as a cause of clinical disorders in reproductive swine herds. Moreover, subsequent studies indicate interspecies transmission and worldwide spreading of PCV3. To date, research related to PCV3 and PCV4 vaccine design is at early stage, and numerous aspects regarding immune response and virus characteristics remain unknown.     

Moving a mountain: Practical insights into mastering a major curriculum reform at a large European medical university

Abstract

Aim: Undergraduate medical education is currently in a fundamental transition towards competency-based programs around the globe. A major curriculum reform implies a dual challenge: the change of the curriculum and the delivering organization. Both are closely interwoven. In this article, we provide practical insights into our approach of managing such a fundamental reform of the large undergraduate medical program at the Charité – Universitätsmedizin Berlin.

Methods: Members of the project management team summarized the key features of the process with reference to the literature.

Results: Starting point was a traditional, discipline-based curriculum that was reformed into a fully integrated, competency-based program. This change process went through three phases: initiation, curriculum development and implementation, and sustainability. We describe from a change management perspective, their main characteristics, and the approaches that were employed to manage them successfully.

Conclusions: Our report is intended to provide practical insights and guidance for those institutions which are yet considering or have already started to undergo a major reform of their undergraduate programs towards competency medical education.     

Synthesis and antiproliferative activity of new mycophenolic acid conjugates with adenosine derivatives

New conjugates of mycophenolic acid (MPA) and adenosine derivatives were synthesized and assessed as potential immunosuppressants on Jurkat cell line and peripheral blood mononuclear cells (PBMC) from healthy donors. As compared to MPA, all compounds were found to be more active against Jurkat cell line. The antiproliferative activities were compared with MPA and adenosine, in both 2′,3′-O-isopropylidene protected and free hydroxyl groups possessing forms. The obtained results were also discussed in terms of selectivity index, defined as SI = IC₅₀/EC₅₀.     

Comparison of Mechanical Properties of Composites Reinforced with Technical Embroidery,UD and Woven Fabric Made of Flax Fibers

The main purpose of the article is to present the possibilities of producing composite reinforcement with the use of a computer embroidery machine. The study below presents the results of strength tests of composites containing technical embroidery, woven fabric, and UD fabric as the reinforcement. Each of the samples was made of the same material—flax roving. The samples differed from each other in the arrangement of layers in the reinforcement. The composites were made using the infusion method with epoxy resin. The embroidery was made on a ZSK embroidery machine, type JCZA 0109-550. A total of 12 types of composites were produced and tested. The test material was subjected to strength tests—tensile strength, tensile elongation, and shear strength, on the INSTRON machine. As the research showed, the use of technical embroidery as a composite reinforcement increases its tensile strength. Furthermore, the use of embroidery is a vertical reinforcement of the composite and prevents the formation of interlayer cracks. The technology of technical embroidery allows for optimizing the mechanical values of the composite reinforcement.     

Role of the Otx1 gene in cell differentiation of mammalian cortex

This study analyses by immunohistochemical methods the effects of the deletion of the Otx1 gene on 12 areas of the cerebral cortex and on neurons expressing Ca-binding proteins (CaBP), such as parvalbumin (Pv) and calbindin-D28K (Cb). We found that the deletion of the Otx1 gene modified differently the various cortical areas. The decrease in cortical thickness ranged from 29.35 to 9.85% and the reduction in cellular population from 35.90 to 3.65% in the different cortical areas. The influence of the Otx1 gene concerns all cortical layers with variable effects on different cortical areas. The cellular population of cerebral cortex considered as a whole was reduced by 20.67%, Pv-positive (Pv+) cells by 58.01% and Cb-positive (Cb+) cells by 51.54%. The quantitative distribution of Pv+ and Cb+ cells varied independently in the different cortical areas. Topographic analysis of CaBP cells in Otx1-null mice (Otx1(-/-)) showed that Pv+ cells were principally distributed in layers IV and V and Cb+ cells in layers V and VI. Given that in the development of wild-type mice both cell types first appear in deep layers and later spread to superficial ones, the segregation of CaBP neurons in inner layers of Otx1(-/-) animals is an index of the immaturity of the cerebral cortex of these animals. This study showed that the Otx1 gene has a more complex role than previously reported, as it is involved in the maturation and differentiation of various cerebral cortices, and, specifically, in the development of CaBP cells.     

A comprehensive approach to identification of pathogenic FANCA variants in Fanconi anemia patients and their families

Fanconi anemia (FA) is a rare recessive DNA repair deficiency resulting from mutations in one of at least 22 genes. Two‐thirds of FA families harbor mutations in FANCA. To genotype patients in the International Fanconi Anemia Registry (IFAR) we employed multiple methodologies, screening 216 families for FANCA mutations. We describe identification of 57 large deletions and 261 sequence variants, in 159 families. All but seven families harbored distinct combinations of two mutations demonstrating high heterogeneity. Pathogenicity of the 18 novel missense variants was analyzed functionally by determining the ability of the mutant cDNA to improve the survival of a FANCA‐null cell line when treated with MMC. Overexpressed pathogenic missense variants were found to reside in the cytoplasm, and nonpathogenic in the nucleus. RNA analysis demonstrated that two variants (c.522G > C and c.1565A > G), predicted to encode missense variants, which were determined to be nonpathogenic by a functional assay, caused skipping of exons 5 and 16, respectively, and are most likely pathogenic. We report 48 novel FANCA sequence variants. Defining both variants in a large patient cohort is a major step toward cataloging all FANCA variants, and permitting studies of genotype–phenotype correlations.     

Thermophilic potentially pathogenic amoebae isolated from natural water bodies in Poland and their molecular characterization

The free-living amoebae (FLA) may live in the environment and also within other organisms as parasites and then they are called amphizoic. They are potentially pathogenic for humans and animals and are found in water that is a source of infection. The aim of this study was molecular detection and identification of these FLA in natural water bodies in North-Western Poland to evaluate the risk of the pathogenic amoebae infections. We examined surface water samples collected from 50 sites and first, the tolerance thermic test was performed in order to select thermophilic, potentially pathogenic strains. For molecular identification of FLA, regions of 18S rDNA, 16S rDNA and intergenic spacers were amplified. Acanthamoeba T4 and T16 genotypes of 18S rDNA gene and 18S rDNA of H. vermiformis were detected. We identified two variants of Acanthamoeba T4 genotype, two variants of Acanthamoeba T16 genotype and one variant of H. vermiformis. Identification of the T16 genotype and H. vermiformis in water was for the first time in Poland. Additionally, we made attempts to adapt the RLB method for detection and differentiation of FLA species and strains. PCR seems to be more sensitive than RLB hybridization, though.     

Establishment of high-resolution FISH mapping system and its application for molecular cytogenetic characterization of chromosomes in newt, <Emphasis Type="Italic">Cynops pyrrhogaster</Emphasis> (Urodela,Amphibia)

Urodele amphibians (newts and salamanders) are important animal models for understanding regeneration mechanisms and genome evolution. We constructed ideograms of BrdU/dT- and C-banded karyotypes in the Japanese fire-belly newt, Cynops pyrrhogaster, which is useful as a model animal with extremely high ability of regeneration. We also established a high-resolution FISH mapping system for newts, and localized satellite DNA sequences, 18S rDNAs, telomeric (TTAGGG)n repeats and seven functional genes, including genes associated with lens regeneration, tyrosinase and two types of gamma crystallins, to chromosomes of the newt. The 18S rDNAs were localized to three chromosomal pairs in males, whereas the chromosomal locations were highly variable in females. No hybridization signals were detected for the telomeric (TTAGGG)n sequence. All three lens regeneration-related genes were mapped on the short arm of chromosome 7, suggesting that the location of the genes in the same linkage group may be correlated with the regulation of gene expression associated with chromatin dynamics in interphase nuclei during regeneration. The chromosomal distribution and nucleotide sequences of pericentric satellite DNA sequences were well conserved between C. pyrrhogaster and European newts; in contrast, there was species specificity of nucleotide sequences for centromere-specific satellite DNAs.     

Median arcuate ligament syndrome: Predictor of ischemic complications?

Median arcuate ligament syndrome (MALS) is a pathologic entity that can affect the celiac axis. Due to the extensive collateral network of mesenteric circulation, stenosis of one mesenteric artery does not lead to significant symptoms. The purpose of this study was to describe multidetector computed tomography (MDCT) angiography findings of celiac artery entrapment by the median arcuate ligament and determine those patients with high risks of ischemic complications. From January 2012 to March 2016, 103 patients with celiac artery (CA) compression by median arcuate ligament were detected. In 23 patients collateral circulation was developed. In order to investigate the problem, we managed to estimate the correlation between range of stenosis of CA and presence of collateral circulation between the celiac artery (CA) and superior mesenteric artery (SMA). A statistically significant correlation was found between range of CA stenosis and collateral circulation presence (Spearman's correlation coefficient 0.339, P < 0.0001). In conclusions, based on our observations, we hypothesize that ischemia as a result of mesenteric vessel narrowing by the median arcuate ligament may occur more often than indicated by clinical symptoms and described in literature. Clin. Anat. 29:1025–1030, 2016. © 2016 Wiley Periodicals, Inc.     

Morphological and cytogenetic analysis of human giant oocytes and giant embryos

BACKGROUND: Giant binuclear oocytes occur with considerable frequency in human ovaries, but their ultimate fate remains unknown. We report the morphology, cytogenetics and developmental potential of human giant oocytes from patients undergoing assisted reproductive technologies. METHODS AND RESULTS: A total of 44 giant oocytes was collected from patients aged 22-44 years old, with an overall frequency of 0.3% (44/14 272 oocytes). Giant oocytes were approximately 30% larger in diameter than normal oocytes (mean 200.4 versus 154.7 micro m, P = 0.0001). Two different morphological patterns were observed among giant unfertilized and fertilized oocytes. All unfertilized oocytes appeared to be diploid and contained either one or two metaphase plates (46 or 2 x 23 chromosomes), and one or two polar bodies respectively. Consequently, fertilized giant oocytes exhibited either two or three pronuclei, or two or four polar bodies. Both types of giant zygotes were capable of normal cleavage and development to blastocyst stage. Four giant embryos were analysed by interphase fluorescence in-situ hybridization using probes for chromosomes 9, 22, X and Y, and all appeared chromosomally abnormal with numerical alterations indicative of ploidy change. CONCLUSIONS: Giant oocytes might be a possible source of human digynic triploidy. To avoid undesired miscarriages, giant embryos originated from either two- or three-pronuclear giant zygotes should be excluded from uterine transfers.